New Techniques for MEA-Based Interrogation of Human

Stem Cell Derived Cardiomyocytes to Support CiPA and

In-vitro Safety Pharmacology Assays

Blake Anson PhD Cellular Dynamics International

Jim Ross PhD Axion Biosystems

Oct 21 2014

Outline

Proarrhythmia Testing

- moving from single channel to holistic

assessments

iCell Cardiomyocytes

Population Diversity

Key manufacturing components

3

In-vitro detection of proarrhythmia

The road to in-vitro

proarrhythmia testinghellip

hellip started in a fly

4

Drug ndashinduced Electrophysiological Aberrations

not a new phenomenon

Quinidine Syncope and Delayed

Repolarization SyndromesReynolds E and Vander Ark C M

Modern Concepts of Cardiovascular Disease 45117-

122 1976

Davies et al BMJ 1989298

Wyosowski and Bacsanyi NEJM 1995335

Astemizole-induced Arrhythmmia

From Vorperian et al JACC 199615

hellipbut took on a new meaning when

caused by non-cardiac compounds

5

Fruit flies provided insight to arrhythmia

Leg shaking

EAG mutant

(ether-a-go-go)

Wild

type

EAG Gene Ether-sedated Drosophila (Fruit Flies)

B Ganetzky

6

hERG is a member of the EAG

superfamily of K+ channels

Library Screen(s)

Hippocampal mRNA

Hit

Analysis

Heterologous

Expression

hERG ndash human ether a

go-go related gene

The hERG gene is linked

to Long QT Syndrome

EAG ndashether a go-go

gene sequence

The hERG gene

encdes lKr

7

DIA

Arrhythmogenic drugs block hERG channels and

prolong the cardiac AP

Zhou and January 1997

8

hERG and In-vitro Safety Assessments

Highly sensitive with

questionable specificity

9

Comprehensive in-vitro Proarrhythmia Testing

The Future

Comprehensive in-vitro Proarrhythmia

Assessment (CiPA)1 Assess effects on multiple individual ion

channels

2 Model effects (if any) on the ventricular

action potential and proarrhythmia

3 Verify conclusions with cardiomyocyte

recordings

httpwwwilsiextraorghesisciencecardiaccipa

Sager et al American Heart Journal (2014)

10

iCell CardiomyocytesDevelopment Regulatory Guidance

Nature Reviews Drug Discovery (Aug Sept 2013)

Product launch regulatory evaluation in 3 years

iPS cell-derived cardiomyocytes are

being evaluated for use in arrhythmia

assessment amp as a replacement for

thorough QT studies

11

Company Overview

Cellular Dynamics International (CDI) is the worldrsquos largest producer of human

iPS cells and iPS cell-derived cell types

Headquartered in Madison WI

Currently employs ~138 total staff

~650 yrs human stem cell experience

gt800 patents (owned or licensed)

Core competencies

Creation and culture of human iPS cells

Normal and disease phenotypes

Genetic engineering of iPS cells

Lineage and pathway-specific markers can be introduced

Development of new differentiation protocols

Differentiated cells from all three germ layers

Manufacture of human iPS cell-derived cell types

Scalable production of highly purified cells

Partnership with iPS Academia Japan enables access

and support for CDIrsquos products in Japan

12

iCell Cardiomyocytes

Human Cardiomyocytes

gt95 pure cryopreserved ready to use

gt4x106 cardiomyocytes per unit

Normal human biology

Broad platform utility for life science research drug

discovery and toxicity testing

Improved workflow with greater predictivity

Full product solution unlimited quantities

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

Outline

Proarrhythmia Testing

- moving from single channel to holistic

assessments

iCell Cardiomyocytes

Population Diversity

Key manufacturing components

3

In-vitro detection of proarrhythmia

The road to in-vitro

proarrhythmia testinghellip

hellip started in a fly

4

Drug ndashinduced Electrophysiological Aberrations

not a new phenomenon

Quinidine Syncope and Delayed

Repolarization SyndromesReynolds E and Vander Ark C M

Modern Concepts of Cardiovascular Disease 45117-

122 1976

Davies et al BMJ 1989298

Wyosowski and Bacsanyi NEJM 1995335

Astemizole-induced Arrhythmmia

From Vorperian et al JACC 199615

hellipbut took on a new meaning when

caused by non-cardiac compounds

5

Fruit flies provided insight to arrhythmia

Leg shaking

EAG mutant

(ether-a-go-go)

Wild

type

EAG Gene Ether-sedated Drosophila (Fruit Flies)

B Ganetzky

6

hERG is a member of the EAG

superfamily of K+ channels

Library Screen(s)

Hippocampal mRNA

Hit

Analysis

Heterologous

Expression

hERG ndash human ether a

go-go related gene

The hERG gene is linked

to Long QT Syndrome

EAG ndashether a go-go

gene sequence

The hERG gene

encdes lKr

7

DIA

Arrhythmogenic drugs block hERG channels and

prolong the cardiac AP

Zhou and January 1997

8

hERG and In-vitro Safety Assessments

Highly sensitive with

questionable specificity

9

Comprehensive in-vitro Proarrhythmia Testing

The Future

Comprehensive in-vitro Proarrhythmia

Assessment (CiPA)1 Assess effects on multiple individual ion

channels

2 Model effects (if any) on the ventricular

action potential and proarrhythmia

3 Verify conclusions with cardiomyocyte

recordings

httpwwwilsiextraorghesisciencecardiaccipa

Sager et al American Heart Journal (2014)

10

iCell CardiomyocytesDevelopment Regulatory Guidance

Nature Reviews Drug Discovery (Aug Sept 2013)

Product launch regulatory evaluation in 3 years

iPS cell-derived cardiomyocytes are

being evaluated for use in arrhythmia

assessment amp as a replacement for

thorough QT studies

11

Company Overview

Cellular Dynamics International (CDI) is the worldrsquos largest producer of human

iPS cells and iPS cell-derived cell types

Headquartered in Madison WI

Currently employs ~138 total staff

~650 yrs human stem cell experience

gt800 patents (owned or licensed)

Core competencies

Creation and culture of human iPS cells

Normal and disease phenotypes

Genetic engineering of iPS cells

Lineage and pathway-specific markers can be introduced

Development of new differentiation protocols

Differentiated cells from all three germ layers

Manufacture of human iPS cell-derived cell types

Scalable production of highly purified cells

Partnership with iPS Academia Japan enables access

and support for CDIrsquos products in Japan

12

iCell Cardiomyocytes

Human Cardiomyocytes

gt95 pure cryopreserved ready to use

gt4x106 cardiomyocytes per unit

Normal human biology

Broad platform utility for life science research drug

discovery and toxicity testing

Improved workflow with greater predictivity

Full product solution unlimited quantities

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

3

In-vitro detection of proarrhythmia

The road to in-vitro

proarrhythmia testinghellip

hellip started in a fly

4

Drug ndashinduced Electrophysiological Aberrations

not a new phenomenon

Quinidine Syncope and Delayed

Repolarization SyndromesReynolds E and Vander Ark C M

Modern Concepts of Cardiovascular Disease 45117-

122 1976

Davies et al BMJ 1989298

Wyosowski and Bacsanyi NEJM 1995335

Astemizole-induced Arrhythmmia

From Vorperian et al JACC 199615

hellipbut took on a new meaning when

caused by non-cardiac compounds

5

Fruit flies provided insight to arrhythmia

Leg shaking

EAG mutant

(ether-a-go-go)

Wild

type

EAG Gene Ether-sedated Drosophila (Fruit Flies)

B Ganetzky

6

hERG is a member of the EAG

superfamily of K+ channels

Library Screen(s)

Hippocampal mRNA

Hit

Analysis

Heterologous

Expression

hERG ndash human ether a

go-go related gene

The hERG gene is linked

to Long QT Syndrome

EAG ndashether a go-go

gene sequence

The hERG gene

encdes lKr

7

DIA

Arrhythmogenic drugs block hERG channels and

prolong the cardiac AP

Zhou and January 1997

8

hERG and In-vitro Safety Assessments

Highly sensitive with

questionable specificity

9

Comprehensive in-vitro Proarrhythmia Testing

The Future

Comprehensive in-vitro Proarrhythmia

Assessment (CiPA)1 Assess effects on multiple individual ion

channels

2 Model effects (if any) on the ventricular

action potential and proarrhythmia

3 Verify conclusions with cardiomyocyte

recordings

httpwwwilsiextraorghesisciencecardiaccipa

Sager et al American Heart Journal (2014)

10

iCell CardiomyocytesDevelopment Regulatory Guidance

Nature Reviews Drug Discovery (Aug Sept 2013)

Product launch regulatory evaluation in 3 years

iPS cell-derived cardiomyocytes are

being evaluated for use in arrhythmia

assessment amp as a replacement for

thorough QT studies

11

Company Overview

Cellular Dynamics International (CDI) is the worldrsquos largest producer of human

iPS cells and iPS cell-derived cell types

Headquartered in Madison WI

Currently employs ~138 total staff

~650 yrs human stem cell experience

gt800 patents (owned or licensed)

Core competencies

Creation and culture of human iPS cells

Normal and disease phenotypes

Genetic engineering of iPS cells

Lineage and pathway-specific markers can be introduced

Development of new differentiation protocols

Differentiated cells from all three germ layers

Manufacture of human iPS cell-derived cell types

Scalable production of highly purified cells

Partnership with iPS Academia Japan enables access

and support for CDIrsquos products in Japan

12

iCell Cardiomyocytes

Human Cardiomyocytes

gt95 pure cryopreserved ready to use

gt4x106 cardiomyocytes per unit

Normal human biology

Broad platform utility for life science research drug

discovery and toxicity testing

Improved workflow with greater predictivity

Full product solution unlimited quantities

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

4

Drug ndashinduced Electrophysiological Aberrations

not a new phenomenon

Quinidine Syncope and Delayed

Repolarization SyndromesReynolds E and Vander Ark C M

Modern Concepts of Cardiovascular Disease 45117-

122 1976

Davies et al BMJ 1989298

Wyosowski and Bacsanyi NEJM 1995335

Astemizole-induced Arrhythmmia

From Vorperian et al JACC 199615

hellipbut took on a new meaning when

caused by non-cardiac compounds

5

Fruit flies provided insight to arrhythmia

Leg shaking

EAG mutant

(ether-a-go-go)

Wild

type

EAG Gene Ether-sedated Drosophila (Fruit Flies)

B Ganetzky

6

hERG is a member of the EAG

superfamily of K+ channels

Library Screen(s)

Hippocampal mRNA

Hit

Analysis

Heterologous

Expression

hERG ndash human ether a

go-go related gene

The hERG gene is linked

to Long QT Syndrome

EAG ndashether a go-go

gene sequence

The hERG gene

encdes lKr

7

DIA

Arrhythmogenic drugs block hERG channels and

prolong the cardiac AP

Zhou and January 1997

8

hERG and In-vitro Safety Assessments

Highly sensitive with

questionable specificity

9

Comprehensive in-vitro Proarrhythmia Testing

The Future

Comprehensive in-vitro Proarrhythmia

Assessment (CiPA)1 Assess effects on multiple individual ion

channels

2 Model effects (if any) on the ventricular

action potential and proarrhythmia

3 Verify conclusions with cardiomyocyte

recordings

httpwwwilsiextraorghesisciencecardiaccipa

Sager et al American Heart Journal (2014)

10

iCell CardiomyocytesDevelopment Regulatory Guidance

Nature Reviews Drug Discovery (Aug Sept 2013)

Product launch regulatory evaluation in 3 years

iPS cell-derived cardiomyocytes are

being evaluated for use in arrhythmia

assessment amp as a replacement for

thorough QT studies

11

Company Overview

Cellular Dynamics International (CDI) is the worldrsquos largest producer of human

iPS cells and iPS cell-derived cell types

Headquartered in Madison WI

Currently employs ~138 total staff

~650 yrs human stem cell experience

gt800 patents (owned or licensed)

Core competencies

Creation and culture of human iPS cells

Normal and disease phenotypes

Genetic engineering of iPS cells

Lineage and pathway-specific markers can be introduced

Development of new differentiation protocols

Differentiated cells from all three germ layers

Manufacture of human iPS cell-derived cell types

Scalable production of highly purified cells

Partnership with iPS Academia Japan enables access

and support for CDIrsquos products in Japan

12

iCell Cardiomyocytes

Human Cardiomyocytes

gt95 pure cryopreserved ready to use

gt4x106 cardiomyocytes per unit

Normal human biology

Broad platform utility for life science research drug

discovery and toxicity testing

Improved workflow with greater predictivity

Full product solution unlimited quantities

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

5

Fruit flies provided insight to arrhythmia

Leg shaking

EAG mutant

(ether-a-go-go)

Wild

type

EAG Gene Ether-sedated Drosophila (Fruit Flies)

B Ganetzky

6

hERG is a member of the EAG

superfamily of K+ channels

Library Screen(s)

Hippocampal mRNA

Hit

Analysis

Heterologous

Expression

hERG ndash human ether a

go-go related gene

The hERG gene is linked

to Long QT Syndrome

EAG ndashether a go-go

gene sequence

The hERG gene

encdes lKr

7

DIA

Arrhythmogenic drugs block hERG channels and

prolong the cardiac AP

Zhou and January 1997

8

hERG and In-vitro Safety Assessments

Highly sensitive with

questionable specificity

9

Comprehensive in-vitro Proarrhythmia Testing

The Future

Comprehensive in-vitro Proarrhythmia

Assessment (CiPA)1 Assess effects on multiple individual ion

channels

2 Model effects (if any) on the ventricular

action potential and proarrhythmia

3 Verify conclusions with cardiomyocyte

recordings

httpwwwilsiextraorghesisciencecardiaccipa

Sager et al American Heart Journal (2014)

10

iCell CardiomyocytesDevelopment Regulatory Guidance

Nature Reviews Drug Discovery (Aug Sept 2013)

Product launch regulatory evaluation in 3 years

iPS cell-derived cardiomyocytes are

being evaluated for use in arrhythmia

assessment amp as a replacement for

thorough QT studies

11

Company Overview

Cellular Dynamics International (CDI) is the worldrsquos largest producer of human

iPS cells and iPS cell-derived cell types

Headquartered in Madison WI

Currently employs ~138 total staff

~650 yrs human stem cell experience

gt800 patents (owned or licensed)

Core competencies

Creation and culture of human iPS cells

Normal and disease phenotypes

Genetic engineering of iPS cells

Lineage and pathway-specific markers can be introduced

Development of new differentiation protocols

Differentiated cells from all three germ layers

Manufacture of human iPS cell-derived cell types

Scalable production of highly purified cells

Partnership with iPS Academia Japan enables access

and support for CDIrsquos products in Japan

12

iCell Cardiomyocytes

Human Cardiomyocytes

gt95 pure cryopreserved ready to use

gt4x106 cardiomyocytes per unit

Normal human biology

Broad platform utility for life science research drug

discovery and toxicity testing

Improved workflow with greater predictivity

Full product solution unlimited quantities

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

6

hERG is a member of the EAG

superfamily of K+ channels

Library Screen(s)

Hippocampal mRNA

Hit

Analysis

Heterologous

Expression

hERG ndash human ether a

go-go related gene

The hERG gene is linked

to Long QT Syndrome

EAG ndashether a go-go

gene sequence

The hERG gene

encdes lKr

7

DIA

Arrhythmogenic drugs block hERG channels and

prolong the cardiac AP

Zhou and January 1997

8

hERG and In-vitro Safety Assessments

Highly sensitive with

questionable specificity

9

Comprehensive in-vitro Proarrhythmia Testing

The Future

Comprehensive in-vitro Proarrhythmia

Assessment (CiPA)1 Assess effects on multiple individual ion

channels

2 Model effects (if any) on the ventricular

action potential and proarrhythmia

3 Verify conclusions with cardiomyocyte

recordings

httpwwwilsiextraorghesisciencecardiaccipa

Sager et al American Heart Journal (2014)

10

iCell CardiomyocytesDevelopment Regulatory Guidance

Nature Reviews Drug Discovery (Aug Sept 2013)

Product launch regulatory evaluation in 3 years

iPS cell-derived cardiomyocytes are

being evaluated for use in arrhythmia

assessment amp as a replacement for

thorough QT studies

11

Company Overview

Cellular Dynamics International (CDI) is the worldrsquos largest producer of human

iPS cells and iPS cell-derived cell types

Headquartered in Madison WI

Currently employs ~138 total staff

~650 yrs human stem cell experience

gt800 patents (owned or licensed)

Core competencies

Creation and culture of human iPS cells

Normal and disease phenotypes

Genetic engineering of iPS cells

Lineage and pathway-specific markers can be introduced

Development of new differentiation protocols

Differentiated cells from all three germ layers

Manufacture of human iPS cell-derived cell types

Scalable production of highly purified cells

Partnership with iPS Academia Japan enables access

and support for CDIrsquos products in Japan

12

iCell Cardiomyocytes

Human Cardiomyocytes

gt95 pure cryopreserved ready to use

gt4x106 cardiomyocytes per unit

Normal human biology

Broad platform utility for life science research drug

discovery and toxicity testing

Improved workflow with greater predictivity

Full product solution unlimited quantities

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

7

DIA

Arrhythmogenic drugs block hERG channels and

prolong the cardiac AP

Zhou and January 1997

8

hERG and In-vitro Safety Assessments

Highly sensitive with

questionable specificity

9

Comprehensive in-vitro Proarrhythmia Testing

The Future

Comprehensive in-vitro Proarrhythmia

Assessment (CiPA)1 Assess effects on multiple individual ion

channels

2 Model effects (if any) on the ventricular

action potential and proarrhythmia

3 Verify conclusions with cardiomyocyte

recordings

httpwwwilsiextraorghesisciencecardiaccipa

Sager et al American Heart Journal (2014)

10

iCell CardiomyocytesDevelopment Regulatory Guidance

Nature Reviews Drug Discovery (Aug Sept 2013)

Product launch regulatory evaluation in 3 years

iPS cell-derived cardiomyocytes are

being evaluated for use in arrhythmia

assessment amp as a replacement for

thorough QT studies

11

Company Overview

Cellular Dynamics International (CDI) is the worldrsquos largest producer of human

iPS cells and iPS cell-derived cell types

Headquartered in Madison WI

Currently employs ~138 total staff

~650 yrs human stem cell experience

gt800 patents (owned or licensed)

Core competencies

Creation and culture of human iPS cells

Normal and disease phenotypes

Genetic engineering of iPS cells

Lineage and pathway-specific markers can be introduced

Development of new differentiation protocols

Differentiated cells from all three germ layers

Manufacture of human iPS cell-derived cell types

Scalable production of highly purified cells

Partnership with iPS Academia Japan enables access

and support for CDIrsquos products in Japan

12

iCell Cardiomyocytes

Human Cardiomyocytes

gt95 pure cryopreserved ready to use

gt4x106 cardiomyocytes per unit

Normal human biology

Broad platform utility for life science research drug

discovery and toxicity testing

Improved workflow with greater predictivity

Full product solution unlimited quantities

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

8

hERG and In-vitro Safety Assessments

Highly sensitive with

questionable specificity

9

Comprehensive in-vitro Proarrhythmia Testing

The Future

Comprehensive in-vitro Proarrhythmia

Assessment (CiPA)1 Assess effects on multiple individual ion

channels

2 Model effects (if any) on the ventricular

action potential and proarrhythmia

3 Verify conclusions with cardiomyocyte

recordings

httpwwwilsiextraorghesisciencecardiaccipa

Sager et al American Heart Journal (2014)

10

iCell CardiomyocytesDevelopment Regulatory Guidance

Nature Reviews Drug Discovery (Aug Sept 2013)

Product launch regulatory evaluation in 3 years

iPS cell-derived cardiomyocytes are

being evaluated for use in arrhythmia

assessment amp as a replacement for

thorough QT studies

11

Company Overview

Cellular Dynamics International (CDI) is the worldrsquos largest producer of human

iPS cells and iPS cell-derived cell types

Headquartered in Madison WI

Currently employs ~138 total staff

~650 yrs human stem cell experience

gt800 patents (owned or licensed)

Core competencies

Creation and culture of human iPS cells

Normal and disease phenotypes

Genetic engineering of iPS cells

Lineage and pathway-specific markers can be introduced

Development of new differentiation protocols

Differentiated cells from all three germ layers

Manufacture of human iPS cell-derived cell types

Scalable production of highly purified cells

Partnership with iPS Academia Japan enables access

and support for CDIrsquos products in Japan

12

iCell Cardiomyocytes

Human Cardiomyocytes

gt95 pure cryopreserved ready to use

gt4x106 cardiomyocytes per unit

Normal human biology

Broad platform utility for life science research drug

discovery and toxicity testing

Improved workflow with greater predictivity

Full product solution unlimited quantities

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

9

Comprehensive in-vitro Proarrhythmia Testing

The Future

Comprehensive in-vitro Proarrhythmia

Assessment (CiPA)1 Assess effects on multiple individual ion

channels

2 Model effects (if any) on the ventricular

action potential and proarrhythmia

3 Verify conclusions with cardiomyocyte

recordings

httpwwwilsiextraorghesisciencecardiaccipa

Sager et al American Heart Journal (2014)

10

iCell CardiomyocytesDevelopment Regulatory Guidance

Nature Reviews Drug Discovery (Aug Sept 2013)

Product launch regulatory evaluation in 3 years

iPS cell-derived cardiomyocytes are

being evaluated for use in arrhythmia

assessment amp as a replacement for

thorough QT studies

11

Company Overview

Cellular Dynamics International (CDI) is the worldrsquos largest producer of human

iPS cells and iPS cell-derived cell types

Headquartered in Madison WI

Currently employs ~138 total staff

~650 yrs human stem cell experience

gt800 patents (owned or licensed)

Core competencies

Creation and culture of human iPS cells

Normal and disease phenotypes

Genetic engineering of iPS cells

Lineage and pathway-specific markers can be introduced

Development of new differentiation protocols

Differentiated cells from all three germ layers

Manufacture of human iPS cell-derived cell types

Scalable production of highly purified cells

Partnership with iPS Academia Japan enables access

and support for CDIrsquos products in Japan

12

iCell Cardiomyocytes

Human Cardiomyocytes

gt95 pure cryopreserved ready to use

gt4x106 cardiomyocytes per unit

Normal human biology

Broad platform utility for life science research drug

discovery and toxicity testing

Improved workflow with greater predictivity

Full product solution unlimited quantities

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

10

iCell CardiomyocytesDevelopment Regulatory Guidance

Nature Reviews Drug Discovery (Aug Sept 2013)

Product launch regulatory evaluation in 3 years

iPS cell-derived cardiomyocytes are

being evaluated for use in arrhythmia

assessment amp as a replacement for

thorough QT studies

11

Company Overview

Cellular Dynamics International (CDI) is the worldrsquos largest producer of human

iPS cells and iPS cell-derived cell types

Headquartered in Madison WI

Currently employs ~138 total staff

~650 yrs human stem cell experience

gt800 patents (owned or licensed)

Core competencies

Creation and culture of human iPS cells

Normal and disease phenotypes

Genetic engineering of iPS cells

Lineage and pathway-specific markers can be introduced

Development of new differentiation protocols

Differentiated cells from all three germ layers

Manufacture of human iPS cell-derived cell types

Scalable production of highly purified cells

Partnership with iPS Academia Japan enables access

and support for CDIrsquos products in Japan

12

iCell Cardiomyocytes

Human Cardiomyocytes

gt95 pure cryopreserved ready to use

gt4x106 cardiomyocytes per unit

Normal human biology

Broad platform utility for life science research drug

discovery and toxicity testing

Improved workflow with greater predictivity

Full product solution unlimited quantities

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

11

Company Overview

Cellular Dynamics International (CDI) is the worldrsquos largest producer of human

iPS cells and iPS cell-derived cell types

Headquartered in Madison WI

Currently employs ~138 total staff

~650 yrs human stem cell experience

gt800 patents (owned or licensed)

Core competencies

Creation and culture of human iPS cells

Normal and disease phenotypes

Genetic engineering of iPS cells

Lineage and pathway-specific markers can be introduced

Development of new differentiation protocols

Differentiated cells from all three germ layers

Manufacture of human iPS cell-derived cell types

Scalable production of highly purified cells

Partnership with iPS Academia Japan enables access

and support for CDIrsquos products in Japan

12

iCell Cardiomyocytes

Human Cardiomyocytes

gt95 pure cryopreserved ready to use

gt4x106 cardiomyocytes per unit

Normal human biology

Broad platform utility for life science research drug

discovery and toxicity testing

Improved workflow with greater predictivity

Full product solution unlimited quantities

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

12

iCell Cardiomyocytes

Human Cardiomyocytes

gt95 pure cryopreserved ready to use

gt4x106 cardiomyocytes per unit

Normal human biology

Broad platform utility for life science research drug

discovery and toxicity testing

Improved workflow with greater predictivity

Full product solution unlimited quantities

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

13

iCell CardiomyocytesCharacterization

Electrophysiology E-C Coupling Contractility

Enables mechanistic toxicity testing

Whole-Genome Gene Expression

Relevant amp stable over time in culture

------ Adult myocardium____ d28 iCell CM____ d120 iCell CM

____ PromoCell____ Celprogen

-------Myocardium

Babiarz et al 2012 Kattman et al 2011 Rana et al 2012 Ma et al 2011

(For a complete list of iCell Cardiomyocytes publications go to

wwwcellulardynamicscom)

Metabolism

Recapitulates normal human

cardiac function

Protein Expression

Appropriate for interrogating

mitochondrial toxicity

iCell Cardiomyocytes native human biology enables

Mechanistic interrogation of cardiac function

Toxicity testing disruption of normal processes

Disease modeling corruption of normal processes

Well represented in the peer reviewed literature

~40 iCell Cardiomyocytes pubs to-date

More than all other commercial iPSC-CMs combined

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

14

CDI CommitmentQuality Quantity Purity

Quality

Quantity Purity

Exhibit key cellular characteristics

Recapitulate normal human biology

Reproducible

Known and relevant genotype

Sufficient to support HTP drug screening

and safety testing

Currently 1Bn iCell Cardiomyocytesday

Ce

ll P

uri

ty

Days in Culture

Target Cell (non proliferating)

Non-Target Cell (proliferating)

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

15

StandardizationManufacturing Benchmarks

NHLBI Next Generation Genetic

Association Studies(RFA-HL-11-066)

250 patient samples - HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes from

all 250 individuals

Induce hypertrophy phenotype perform

molecular analyses

Correlate GWAS findings with in vitro

phenotype

Scale-Out

Manufacturing

bull 1000rsquos of individuals

bull Billions of cells

Scale-Up

Manufacturing

bull Quality

bull Quantity

bull Purity

CDI Manufacturing Benchmarks (cells per day gt95 purity)

2 billion iPS cells

1 billion cardiomyocytes

1 billion neurons

05 billion endothelial cells

04 billion hepatocytes

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

16

NHLBI Next Generation Genetic

Association Studies (RFA-HL-11-066)

250 patient samples ndash HyperGEN cohort

GWAS ndash Left Ventricular Hypertrophy (LVH)

Derive iPS cells and cardiomyocytes

Induce hypertrophy perform molecular analyses

Correlate GWAS findings with in vitro phenotype

Progress as of July 2014

250 donors reprogrammed

Differentiation protocol optimized to work robustly across all lines

128 iPS cell lines (1 per donor) are differentiated or in progress

Cardiomyocytes from 89 donors cryopreserved amp all pass QC

20 batches of cardiomyocytes are in currently being tested in

hypertrophy assays

Initial data show Et-1 EC50 correlation with progression of disease (Uli Broeckel MCOW)

Progress ReportPopulation genomics and left ventricular hypertrophy

CDIrsquos iPSC technology is enabling population studies

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

17

CIRM AwardiPS Cell Manufacture amp Banking

California Institute for Regenerative Medicine (CIRM)

Human iPS Cell Initiative ndash 3 Awards

Sample Collection iPSC Derivation (CDI) iPS Cell Banking

iPS Cell Derivation (CDI)

3000 donors (healthy amp disease phenotypes)

3 iPS cell clones per donor

Disease categories epilepsy autism cerebral palsy cardiomyopathy Alzheimerrsquos

disease eye diseases hepatitis (HCV) non-alcoholic steatohepatitis (NASH)

pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts

Episomal ldquofootprint-freerdquo method

CDI ndash Coriell Partnership

Extensive collaboration to bring together expertise in electronic record-keeping

sample tracking iPS cell derivation amp characterization cell banking amp distribution

Joint facility located within the Buck Institute Novato CA

Demonstrated success in generating high quality iPSC lsquopopulationsrsquo

Well poised to enable population diversity in toxicity and safety assessments

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

QMS Framework Overview

Key Systems Objectives

QAQC Compliance and product consistency

Standard Operating Procedures Consistent procedures

CalibrationQualVal Equipmentfacilitiesprocesses fit for intended use

Change Management Changes are documented assessed for risk and tested

CAPA Report correct and prevent product quality issues

Supplier Qual amp Mgmt Quality and reliability of raw materials

Materials Management Control trace and monitor stock inventory

Training Education and proficiency

Complaint Handling Customer satisfaction and continuous improvement

New Product Introduction Improve likelihood that product meets market need

An ISO GMP hybrid QMS system ensures customer safety and satisfaction

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

20

1 Nakamura Y1 Matsuo J (2014) Assessment of testing methods for

drug-induced repolarization delay and arrhythmias in an iPS cell-

derived cardiomyocyte sheet multi-site validation study J

Pharmacol Sci 124(4)494-501

2 Eldridge S Guo L et al (2014) Examining the Protective Role of

ErbB2 Modulation in Human Induced Pluripotent Stem Cell-

Derived Cardiomyocytes Toxicol Sci 2014 Jul 23 pii kfu150

[Epub ahead of print]

3 Kolaja K (2014) Stem cells and stem cell-derived tissues and their

use in safety assessment J Biol Chem 2014 Feb 21289(8)4555-

61

4 Uesugi M Ojima A et al (2014) Low-density plating is sufficient to

induce cardiac hypertrophy and electrical remodeling in highly

purified human iPS cell-derived cardiomyocytes J Pharmacol

Toxicol Methods 69(2)177-88

5 Cameron BJ Gerry AB et al (2013) Identification of a Titin-

derived HLA-A1-presented peptide as a cross-reactive target for

engineered MAGE A3-directed T cells Sci Transl Med

5(197)197ra103

6 Carlson C Koonce C et al (2013) Phenotypic screening with

human iPS cell-derived cardiomyocytes HTS-compatible assays

for interrogating cardiac hypertrophy J Biomol Screen

18(10)1203-11

7 Doherty K Wappel R et al (2013) Multiparameter in vitro toxicity

testing of crizotinib sunitinib erlotinib and nilotinib in human

cardiomyocytes Toxicol Appl Pharmacol 272(1)245-55

8 Fine M Lu F et al (2013) Human Induced Pluripotent Stem Cell-

derived Cardiomyocytes for Studies of Cardiac Ion Transporters

Am J Physiol Cell Physiol 305(5)C481-91

9 Guo L Coyle l et al (2013) Refining the Human iPSC-

Cardiomyocyte Arrhythmic Risk Assessment Model Toxicol Sci

136(2)581-94

10 Harris K Aylott M et al (2013) Comparison of

Electrophysiological Data from Human Induced Pluripotent Stem

Cell Derived Cardiomyoyctes (hiPSC-CMs) to Functional Pre-

clinical Safety Assays Toxicol Sci 134(2)412-26

11 Ivashchenko CY1 Pipes GC et al (2013) Human-induced

pluripotent stem cell-derived cardiomyocytes exhibit temporal

changes in phenotype Am J Physiol Heart Circ Physiol

305(6)H913-22

12 Jehle J Ficker E et al (2013) Mechanisms of Zolpidem-induced

Long QT Ayndrome Acute Inhibition of Recombinant hERG K+

Channels and Action Potential Prolongation in Human

Cardiomyocytes Derived from Induced Pluripotent Stem Cells

British J Pharm 1681215-29

13 Puppala D Collis LP et al (2013) Comparative Gene Expression

Profiling in Human Induced Pluripotent Stem Cell Derived

Cardiocytes and Human and Cynomolgus Heart Tissue Toxicol

Sci 131292-301

14 Rao C Prodromakis T et al (2013) The effect of microgrooved

culture substrates on calcium cycling of cardiac myocytes

derived from human induced pluripotent stem cells Biomaterials

34(10)2399-411

15 Schweikart K Guo L et al (2013) The Effects of Jaspamide on

Human Cardiomyocyte Function and Cardiac Ion Channel Activity

Toxicol in Vitro 27745-51

16 Sirenko O Crittenden C et al (2013) Multiparameter In Vitro

Assessment of Compound Effects on Cardiomyocyte Physiology

Using iPS Cells J Biomol Screening 1839-53

17 Sirenko O Cromwell EF et al (2013) Assessment of beating

parameters in human induced pluripotent stem cells enables

quantitative in vitro screening for cardiotoxicity Toxicol Appl

Pharmacol 273(3)500-07

18 Babiarz JE Ravon M et al (2012) Determination of the Human

Cardiomyocyte mRNA and miRNA Differentiation Network by

Fine-scale Profiling Stem Cells Dev 211956-65

19 Cerignoli R Charlot D et al (2012) High Throughput Measurement

of Ca2+ Dynamics for Drug Risk Assessment in Human Stem Cell-

derived Cardiomyocytes by Kinetic Image Cytometry

J Pharmacol Toxicol Methods 66246-256

20 Lee P Kloss M et al (2012) Simultaneous Voltage and Calcium

Mapping of Genetically Purified Human Induced Pluripotent Stem

Cell-derived Cardiac Myocyte Monolayers Circ Res 1101556-63

21 Mioulane M Foldes G et al (2012) Development of High Content

Imaging Methods for Cell Death Detection in Human Pluripotent

Stem Cell-derived Cardiomyocytes J of Cardiovasc Trans Res

5593-604

22 Rana P Anson BD et al (2012) Characterization of Human-

induced Pluripotent Stem Cell-derived Cardiomyocytes

Bioenergetics and Utilization in Safety Screening Toxicol Sci

130117-31

23 Reynolds JG Geretti E et al (2012) HER2-targeted Liposomal

Doxorubicin Displays Enhanced Anti-tumorigenic Effects without

Associated Cardiotoxicity Toxicol Appl Pharmacol 2621-10

24 Wei H Zhang G et al (2012) Hydrogen Sulfide Suppresses

Outward Rectifier Potassium Currents in Human Pluripotent Stem

Cell-Derived Cardiomyocytes Plos One 7(11)e50641

25 Zhi D Irvin MR et al (2012) Whole-exome Sequencing and an

iPSC-derived Cardiomyocyte Model Provides a Powerful Platform

for Gene Discovery in Left Ventricular Hypertrophy Frontiers in

Genetics 392

26 Cohen JD Babiarz JE et al (2011) Use of Human Stem Cell-

derived Cardiomyocytes to Examine Sunitinib Mediated

Cardiotoxicity and Electrophysiological Alterations Toxicol Appl

Pharmacol 25774-83

27 Guo L Qian JY et al (2011) The Electrophysiological Effects of

Cardiac Glycosides in Human iPSC-derived Cardiomyocytes and

in Guinea Pig Isolated Hearts Cell Physiol Biochem 27453-462

28 Guo L Abrams RM et al (2011) Estimating the Risk of Drug-

induced Proarrhythmia Using Human Induced Pluripotent Stem

Cell-derived Cardiomyocytes Toxicol Sci 123281-289

29 Jonsson MKB Wang QD et al (2011) Impedance-based Detection

of Beating Rhythm and Proarrhythmic Effects of Compounds on

Stem Cell-derived Cardiomyocytes Assay and Drug Dev Tech 91-

11

30 Ma J Guo L et al (2011) High Purity Human-induced Pluripotent

Stem Cell-derived Cardiomyocytes Electrophysiological

Properties of Action Potentials and Ionic Currents Am J Physiol

Heart Circ Physiol 301H2006-H2017

iCell CardiomyocytesMarket Validation (82014)

~40 Peer-reviewed

Publications (102014)

bull Characterization

bull Toxicity testing

bull Disease modeling

21

Measuring Cardiomyocyte FunctionElectrical and Mechanical Activity

iCell Cardiomyocytes provide a human system for

measuring all aspects of EC functionality

22

Predictivity ScreensElectrical Measurements

iPSC within frac12 log of wedge in some cases more sensitive

ldquoMEA assays using iPSC-CMs offer a reliable

cost effective surrogate to preclinical in vitro

testing in addition to the 3Rs (refine reduce

and replace animals in research) benefitrdquoHarris et al Toxicol Sci 2013

23

Summary

Proarrhythmia Testing

- moving toward a cellular mechanistic approach that may take advantage of stem cell cardiomyocytes

iCell Cardiomyocytes

- Human biology validated in the peer-reviewed literature

Population Diversity

- Well poised to enable population testing in toxicity and safety assessments

Key manufacturing components

- Quality is king

New techniques for MEA-based assays to support CiPA and in-vitro

safety pharmacology assays

October 21 2014

Jim Ross PhD CTO Axion Biosystems

1 Introduc+on13 to13 Axion13 2 Brief13 MEA13 Technology13 Review13 3 Role13 of13 MEAs13 in13 CiPA13 4 Arrhythmia13 assessment13 5 Cardiomyocyte13 Pacing13 6 New13 developments13

Agenda

Analysis13

Quality13 Control13

Experiments13

Questions

Answers

Axionrsquos Objective Enable Great Science Complicated Network Electrophysiology Made Accessible

Engineering

Applications

Hardw

areDesign

App

lica1

onsDevelop

men

t

SensorMicrofab

SowareD

esign

Applica1ons Engineering

Electronics

Sensors

Software

Applications Driven Innovation High Performance Scalable Technologies that are Simple to Use

Proarrhythmia Score

Mechanism Based Continuous Scale

Rank Ordered Contextual Data

CiPA Objective

Clinical Assessment Human ECG

Ion Channel Panel

In Silico Simulations

Integrated Human Cellular Studies

Integrated Human Cellular Studies

6

Ion Channel Panel

In Silico Simulations

Clinical Assessment Human ECG

1  Repolarization13

2  Arrhythmia13

3  ScalabilityReproducibility13

4  Risk Assessment13

Myocyte Objective

7

CM-MEA bridges the gap between AP and ECG

Na13 Ca13 K13

Clinical13 ECG13

Ac8on13 Poten8al13

CM-shy‐MEA13 Field13

Poten8al13

813

Criteria13 for13 monitoring13 cardiac13 networks13

(1) Real-shy‐+me13 measurement13 of13 the13 phenotypic13 signal13 voltage13 13

(2) Sufficient13 resolu+on13 to13 capture13 field13 poten+als13

(3) Mul+ple13 recording13 sites13 to13 improve13 reliability13 and13 assess13 field13 poten+al13 propaga+on13 13

(4) Label-shy‐free13 non-shy‐invasive13 opera+on13 to13 observe13 natural13 cell13 func+on13 (5) Preserva+on13 of13 cellular13 interconnec+vity13

A13 grid13 of13 microelectrodes13 interfaces13 with13 electro-shy‐ac+ve13 +ssue13 modeling13 complex13

human13 systems13 in13 a13 dish13

CM-shy‐MEA13 Assay13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

22

Predictivity ScreensElectrical Measurements

iPSC within frac12 log of wedge in some cases more sensitive

ldquoMEA assays using iPSC-CMs offer a reliable

cost effective surrogate to preclinical in vitro

testing in addition to the 3Rs (refine reduce

and replace animals in research) benefitrdquoHarris et al Toxicol Sci 2013

23

Summary

Proarrhythmia Testing

- moving toward a cellular mechanistic approach that may take advantage of stem cell cardiomyocytes

iCell Cardiomyocytes

- Human biology validated in the peer-reviewed literature

Population Diversity

- Well poised to enable population testing in toxicity and safety assessments

Key manufacturing components

- Quality is king

New techniques for MEA-based assays to support CiPA and in-vitro

safety pharmacology assays

October 21 2014

Jim Ross PhD CTO Axion Biosystems

1 Introduc+on13 to13 Axion13 2 Brief13 MEA13 Technology13 Review13 3 Role13 of13 MEAs13 in13 CiPA13 4 Arrhythmia13 assessment13 5 Cardiomyocyte13 Pacing13 6 New13 developments13

Agenda

Analysis13

Quality13 Control13

Experiments13

Questions

Answers

Axionrsquos Objective Enable Great Science Complicated Network Electrophysiology Made Accessible

Engineering

Applications

Hardw

areDesign

App

lica1

onsDevelop

men

t

SensorMicrofab

SowareD

esign

Applica1ons Engineering

Electronics

Sensors

Software

Applications Driven Innovation High Performance Scalable Technologies that are Simple to Use

Proarrhythmia Score

Mechanism Based Continuous Scale

Rank Ordered Contextual Data

CiPA Objective

Clinical Assessment Human ECG

Ion Channel Panel

In Silico Simulations

Integrated Human Cellular Studies

Integrated Human Cellular Studies

6

Ion Channel Panel

In Silico Simulations

Clinical Assessment Human ECG

1  Repolarization13

2  Arrhythmia13

3  ScalabilityReproducibility13

4  Risk Assessment13

Myocyte Objective

7

CM-MEA bridges the gap between AP and ECG

Na13 Ca13 K13

Clinical13 ECG13

Ac8on13 Poten8al13

CM-shy‐MEA13 Field13

Poten8al13

813

Criteria13 for13 monitoring13 cardiac13 networks13

(1) Real-shy‐+me13 measurement13 of13 the13 phenotypic13 signal13 voltage13 13

(2) Sufficient13 resolu+on13 to13 capture13 field13 poten+als13

(3) Mul+ple13 recording13 sites13 to13 improve13 reliability13 and13 assess13 field13 poten+al13 propaga+on13 13

(4) Label-shy‐free13 non-shy‐invasive13 opera+on13 to13 observe13 natural13 cell13 func+on13 (5) Preserva+on13 of13 cellular13 interconnec+vity13

A13 grid13 of13 microelectrodes13 interfaces13 with13 electro-shy‐ac+ve13 +ssue13 modeling13 complex13

human13 systems13 in13 a13 dish13

CM-shy‐MEA13 Assay13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

23

Summary

Proarrhythmia Testing

- moving toward a cellular mechanistic approach that may take advantage of stem cell cardiomyocytes

iCell Cardiomyocytes

- Human biology validated in the peer-reviewed literature

Population Diversity

- Well poised to enable population testing in toxicity and safety assessments

Key manufacturing components

- Quality is king

New techniques for MEA-based assays to support CiPA and in-vitro

safety pharmacology assays

October 21 2014

Jim Ross PhD CTO Axion Biosystems

1 Introduc+on13 to13 Axion13 2 Brief13 MEA13 Technology13 Review13 3 Role13 of13 MEAs13 in13 CiPA13 4 Arrhythmia13 assessment13 5 Cardiomyocyte13 Pacing13 6 New13 developments13

Agenda

Analysis13

Quality13 Control13

Experiments13

Questions

Answers

Axionrsquos Objective Enable Great Science Complicated Network Electrophysiology Made Accessible

Engineering

Applications

Hardw

areDesign

App

lica1

onsDevelop

men

t

SensorMicrofab

SowareD

esign

Applica1ons Engineering

Electronics

Sensors

Software

Applications Driven Innovation High Performance Scalable Technologies that are Simple to Use

Proarrhythmia Score

Mechanism Based Continuous Scale

Rank Ordered Contextual Data

CiPA Objective

Clinical Assessment Human ECG

Ion Channel Panel

In Silico Simulations

Integrated Human Cellular Studies

Integrated Human Cellular Studies

6

Ion Channel Panel

In Silico Simulations

Clinical Assessment Human ECG

1  Repolarization13

2  Arrhythmia13

3  ScalabilityReproducibility13

4  Risk Assessment13

Myocyte Objective

7

CM-MEA bridges the gap between AP and ECG

Na13 Ca13 K13

Clinical13 ECG13

Ac8on13 Poten8al13

CM-shy‐MEA13 Field13

Poten8al13

813

Criteria13 for13 monitoring13 cardiac13 networks13

(1) Real-shy‐+me13 measurement13 of13 the13 phenotypic13 signal13 voltage13 13

(2) Sufficient13 resolu+on13 to13 capture13 field13 poten+als13

(3) Mul+ple13 recording13 sites13 to13 improve13 reliability13 and13 assess13 field13 poten+al13 propaga+on13 13

(4) Label-shy‐free13 non-shy‐invasive13 opera+on13 to13 observe13 natural13 cell13 func+on13 (5) Preserva+on13 of13 cellular13 interconnec+vity13

A13 grid13 of13 microelectrodes13 interfaces13 with13 electro-shy‐ac+ve13 +ssue13 modeling13 complex13

human13 systems13 in13 a13 dish13

CM-shy‐MEA13 Assay13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

New techniques for MEA-based assays to support CiPA and in-vitro

safety pharmacology assays

October 21 2014

Jim Ross PhD CTO Axion Biosystems

1 Introduc+on13 to13 Axion13 2 Brief13 MEA13 Technology13 Review13 3 Role13 of13 MEAs13 in13 CiPA13 4 Arrhythmia13 assessment13 5 Cardiomyocyte13 Pacing13 6 New13 developments13

Agenda

Analysis13

Quality13 Control13

Experiments13

Questions

Answers

Axionrsquos Objective Enable Great Science Complicated Network Electrophysiology Made Accessible

Engineering

Applications

Hardw

areDesign

App

lica1

onsDevelop

men

t

SensorMicrofab

SowareD

esign

Applica1ons Engineering

Electronics

Sensors

Software

Applications Driven Innovation High Performance Scalable Technologies that are Simple to Use

Proarrhythmia Score

Mechanism Based Continuous Scale

Rank Ordered Contextual Data

CiPA Objective

Clinical Assessment Human ECG

Ion Channel Panel

In Silico Simulations

Integrated Human Cellular Studies

Integrated Human Cellular Studies

6

Ion Channel Panel

In Silico Simulations

Clinical Assessment Human ECG

1  Repolarization13

2  Arrhythmia13

3  ScalabilityReproducibility13

4  Risk Assessment13

Myocyte Objective

7

CM-MEA bridges the gap between AP and ECG

Na13 Ca13 K13

Clinical13 ECG13

Ac8on13 Poten8al13

CM-shy‐MEA13 Field13

Poten8al13

813

Criteria13 for13 monitoring13 cardiac13 networks13

(1) Real-shy‐+me13 measurement13 of13 the13 phenotypic13 signal13 voltage13 13

(2) Sufficient13 resolu+on13 to13 capture13 field13 poten+als13

(3) Mul+ple13 recording13 sites13 to13 improve13 reliability13 and13 assess13 field13 poten+al13 propaga+on13 13

(4) Label-shy‐free13 non-shy‐invasive13 opera+on13 to13 observe13 natural13 cell13 func+on13 (5) Preserva+on13 of13 cellular13 interconnec+vity13

A13 grid13 of13 microelectrodes13 interfaces13 with13 electro-shy‐ac+ve13 +ssue13 modeling13 complex13

human13 systems13 in13 a13 dish13

CM-shy‐MEA13 Assay13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

1 Introduc+on13 to13 Axion13 2 Brief13 MEA13 Technology13 Review13 3 Role13 of13 MEAs13 in13 CiPA13 4 Arrhythmia13 assessment13 5 Cardiomyocyte13 Pacing13 6 New13 developments13

Agenda

Analysis13

Quality13 Control13

Experiments13

Questions

Answers

Axionrsquos Objective Enable Great Science Complicated Network Electrophysiology Made Accessible

Engineering

Applications

Hardw

areDesign

App

lica1

onsDevelop

men

t

SensorMicrofab

SowareD

esign

Applica1ons Engineering

Electronics

Sensors

Software

Applications Driven Innovation High Performance Scalable Technologies that are Simple to Use

Proarrhythmia Score

Mechanism Based Continuous Scale

Rank Ordered Contextual Data

CiPA Objective

Clinical Assessment Human ECG

Ion Channel Panel

In Silico Simulations

Integrated Human Cellular Studies

Integrated Human Cellular Studies

6

Ion Channel Panel

In Silico Simulations

Clinical Assessment Human ECG

1  Repolarization13

2  Arrhythmia13

3  ScalabilityReproducibility13

4  Risk Assessment13

Myocyte Objective

7

CM-MEA bridges the gap between AP and ECG

Na13 Ca13 K13

Clinical13 ECG13

Ac8on13 Poten8al13

CM-shy‐MEA13 Field13

Poten8al13

813

Criteria13 for13 monitoring13 cardiac13 networks13

(1) Real-shy‐+me13 measurement13 of13 the13 phenotypic13 signal13 voltage13 13

(2) Sufficient13 resolu+on13 to13 capture13 field13 poten+als13

(3) Mul+ple13 recording13 sites13 to13 improve13 reliability13 and13 assess13 field13 poten+al13 propaga+on13 13

(4) Label-shy‐free13 non-shy‐invasive13 opera+on13 to13 observe13 natural13 cell13 func+on13 (5) Preserva+on13 of13 cellular13 interconnec+vity13

A13 grid13 of13 microelectrodes13 interfaces13 with13 electro-shy‐ac+ve13 +ssue13 modeling13 complex13

human13 systems13 in13 a13 dish13

CM-shy‐MEA13 Assay13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Analysis13

Quality13 Control13

Experiments13

Questions

Answers

Axionrsquos Objective Enable Great Science Complicated Network Electrophysiology Made Accessible

Engineering

Applications

Hardw

areDesign

App

lica1

onsDevelop

men

t

SensorMicrofab

SowareD

esign

Applica1ons Engineering

Electronics

Sensors

Software

Applications Driven Innovation High Performance Scalable Technologies that are Simple to Use

Proarrhythmia Score

Mechanism Based Continuous Scale

Rank Ordered Contextual Data

CiPA Objective

Clinical Assessment Human ECG

Ion Channel Panel

In Silico Simulations

Integrated Human Cellular Studies

Integrated Human Cellular Studies

6

Ion Channel Panel

In Silico Simulations

Clinical Assessment Human ECG

1  Repolarization13

2  Arrhythmia13

3  ScalabilityReproducibility13

4  Risk Assessment13

Myocyte Objective

7

CM-MEA bridges the gap between AP and ECG

Na13 Ca13 K13

Clinical13 ECG13

Ac8on13 Poten8al13

CM-shy‐MEA13 Field13

Poten8al13

813

Criteria13 for13 monitoring13 cardiac13 networks13

(1) Real-shy‐+me13 measurement13 of13 the13 phenotypic13 signal13 voltage13 13

(2) Sufficient13 resolu+on13 to13 capture13 field13 poten+als13

(3) Mul+ple13 recording13 sites13 to13 improve13 reliability13 and13 assess13 field13 poten+al13 propaga+on13 13

(4) Label-shy‐free13 non-shy‐invasive13 opera+on13 to13 observe13 natural13 cell13 func+on13 (5) Preserva+on13 of13 cellular13 interconnec+vity13

A13 grid13 of13 microelectrodes13 interfaces13 with13 electro-shy‐ac+ve13 +ssue13 modeling13 complex13

human13 systems13 in13 a13 dish13

CM-shy‐MEA13 Assay13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Engineering

Applications

Hardw

areDesign

App

lica1

onsDevelop

men

t

SensorMicrofab

SowareD

esign

Applica1ons Engineering

Electronics

Sensors

Software

Applications Driven Innovation High Performance Scalable Technologies that are Simple to Use

Proarrhythmia Score

Mechanism Based Continuous Scale

Rank Ordered Contextual Data

CiPA Objective

Clinical Assessment Human ECG

Ion Channel Panel

In Silico Simulations

Integrated Human Cellular Studies

Integrated Human Cellular Studies

6

Ion Channel Panel

In Silico Simulations

Clinical Assessment Human ECG

1  Repolarization13

2  Arrhythmia13

3  ScalabilityReproducibility13

4  Risk Assessment13

Myocyte Objective

7

CM-MEA bridges the gap between AP and ECG

Na13 Ca13 K13

Clinical13 ECG13

Ac8on13 Poten8al13

CM-shy‐MEA13 Field13

Poten8al13

813

Criteria13 for13 monitoring13 cardiac13 networks13

(1) Real-shy‐+me13 measurement13 of13 the13 phenotypic13 signal13 voltage13 13

(2) Sufficient13 resolu+on13 to13 capture13 field13 poten+als13

(3) Mul+ple13 recording13 sites13 to13 improve13 reliability13 and13 assess13 field13 poten+al13 propaga+on13 13

(4) Label-shy‐free13 non-shy‐invasive13 opera+on13 to13 observe13 natural13 cell13 func+on13 (5) Preserva+on13 of13 cellular13 interconnec+vity13

A13 grid13 of13 microelectrodes13 interfaces13 with13 electro-shy‐ac+ve13 +ssue13 modeling13 complex13

human13 systems13 in13 a13 dish13

CM-shy‐MEA13 Assay13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Proarrhythmia Score

Mechanism Based Continuous Scale

Rank Ordered Contextual Data

CiPA Objective

Clinical Assessment Human ECG

Ion Channel Panel

In Silico Simulations

Integrated Human Cellular Studies

Integrated Human Cellular Studies

6

Ion Channel Panel

In Silico Simulations

Clinical Assessment Human ECG

1  Repolarization13

2  Arrhythmia13

3  ScalabilityReproducibility13

4  Risk Assessment13

Myocyte Objective

7

CM-MEA bridges the gap between AP and ECG

Na13 Ca13 K13

Clinical13 ECG13

Ac8on13 Poten8al13

CM-shy‐MEA13 Field13

Poten8al13

813

Criteria13 for13 monitoring13 cardiac13 networks13

(1) Real-shy‐+me13 measurement13 of13 the13 phenotypic13 signal13 voltage13 13

(2) Sufficient13 resolu+on13 to13 capture13 field13 poten+als13

(3) Mul+ple13 recording13 sites13 to13 improve13 reliability13 and13 assess13 field13 poten+al13 propaga+on13 13

(4) Label-shy‐free13 non-shy‐invasive13 opera+on13 to13 observe13 natural13 cell13 func+on13 (5) Preserva+on13 of13 cellular13 interconnec+vity13

A13 grid13 of13 microelectrodes13 interfaces13 with13 electro-shy‐ac+ve13 +ssue13 modeling13 complex13

human13 systems13 in13 a13 dish13

CM-shy‐MEA13 Assay13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Integrated Human Cellular Studies

6

Ion Channel Panel

In Silico Simulations

Clinical Assessment Human ECG

1  Repolarization13

2  Arrhythmia13

3  ScalabilityReproducibility13

4  Risk Assessment13

Myocyte Objective

7

CM-MEA bridges the gap between AP and ECG

Na13 Ca13 K13

Clinical13 ECG13

Ac8on13 Poten8al13

CM-shy‐MEA13 Field13

Poten8al13

813

Criteria13 for13 monitoring13 cardiac13 networks13

(1) Real-shy‐+me13 measurement13 of13 the13 phenotypic13 signal13 voltage13 13

(2) Sufficient13 resolu+on13 to13 capture13 field13 poten+als13

(3) Mul+ple13 recording13 sites13 to13 improve13 reliability13 and13 assess13 field13 poten+al13 propaga+on13 13

(4) Label-shy‐free13 non-shy‐invasive13 opera+on13 to13 observe13 natural13 cell13 func+on13 (5) Preserva+on13 of13 cellular13 interconnec+vity13

A13 grid13 of13 microelectrodes13 interfaces13 with13 electro-shy‐ac+ve13 +ssue13 modeling13 complex13

human13 systems13 in13 a13 dish13

CM-shy‐MEA13 Assay13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

7

CM-MEA bridges the gap between AP and ECG

Na13 Ca13 K13

Clinical13 ECG13

Ac8on13 Poten8al13

CM-shy‐MEA13 Field13

Poten8al13

813

Criteria13 for13 monitoring13 cardiac13 networks13

(1) Real-shy‐+me13 measurement13 of13 the13 phenotypic13 signal13 voltage13 13

(2) Sufficient13 resolu+on13 to13 capture13 field13 poten+als13

(3) Mul+ple13 recording13 sites13 to13 improve13 reliability13 and13 assess13 field13 poten+al13 propaga+on13 13

(4) Label-shy‐free13 non-shy‐invasive13 opera+on13 to13 observe13 natural13 cell13 func+on13 (5) Preserva+on13 of13 cellular13 interconnec+vity13

A13 grid13 of13 microelectrodes13 interfaces13 with13 electro-shy‐ac+ve13 +ssue13 modeling13 complex13

human13 systems13 in13 a13 dish13

CM-shy‐MEA13 Assay13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

813

Criteria13 for13 monitoring13 cardiac13 networks13

(1) Real-shy‐+me13 measurement13 of13 the13 phenotypic13 signal13 voltage13 13

(2) Sufficient13 resolu+on13 to13 capture13 field13 poten+als13

(3) Mul+ple13 recording13 sites13 to13 improve13 reliability13 and13 assess13 field13 poten+al13 propaga+on13 13

(4) Label-shy‐free13 non-shy‐invasive13 opera+on13 to13 observe13 natural13 cell13 func+on13 (5) Preserva+on13 of13 cellular13 interconnec+vity13

A13 grid13 of13 microelectrodes13 interfaces13 with13 electro-shy‐ac+ve13 +ssue13 modeling13 complex13

human13 systems13 in13 a13 dish13

CM-shy‐MEA13 Assay13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

A13 grid13 of13 microelectrodes13 interfaces13 with13 electro-shy‐ac+ve13 +ssue13 modeling13 complex13

human13 systems13 in13 a13 dish13

CM-shy‐MEA13 Assay13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Measures13 13 Field13 poten+al13 dura+on13 (ldquoQTrdquo)13 Conduc+on13 velocity13 Beat13 rate13 Field13 poten+al13 metrics13 (Amplitude13 Slope13 etc)13

Applica+ons13 13 Cardiac13 Safety13 Screening13 Drug13 Discovery13 Func+onal13 Disease13 Models13 Stem13 Cells13 Pa+ent-shy‐specific13 Therapies13 13 13

Propagation

100micro

V

1 sec15 2 25 3 35 4 45 5 55

minus1

0

1

2

3

x 10minus4

Depolarization

ms

0

1

2

3

4 Propagation Delay

Repolarization

CM-shy‐MEA13 Assay13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

MEAs in CiPA Current Status Characterizing cross-site reliability of CM Assays

1113

13 MEA13 Pilot13 Studies13 beginning13 in13 October13 201413 13

1213 Pilot13 Sites13 including13 Cyprotex13 Bristol-shy‐Myers13 Squibb13 Chantest13 NCI13 Janssen13 (JNJ)13 Merck13 NMI13 Sanofi13 and13 others13

1113 of13 1213 test13 sites13 are13 independent13 from13 the13 MEA13 supplier13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Tracking Repolarization Direct Measures of Depolarization amp Repolarization

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Tracking Repolarization Assessing stable beating

Con+nuous13 monitoring13 enables13 evalua+on13 of13 stable13 spontaneous13 bea+ng13 Red13 data13 points13 highlight13 the13 most13 stable13 region13 of13 spontaneous13 bea+ng13 between13 1513 ndash13 2013 minutes13 post13 dose13 avoiding13 a13 dri^13 in13 beat13 rate13 at13 1813 minutes13 (red13 square)13 13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Dofe+lide13

Quinidine13

Detecting Arrhythmia Beat Irregularity

Beat13 irregulari+es13 can13 emerge13 and13 disappear13 over13 +me13 13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Beat13 Irregularity13 NotchEAD13 Alternans13 amp13 Triggered13 Ac+vity13

Quinidine 1microM

Nakamura13 201413

Navarrete13 201313

Time13 (sec)13

BP13 (sec)13

Harris13 201313

Detecting Arrhythmia Arrhythmic Event Classification

MEA13 classifies13 aberrant13 events13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

DMSO13 113 microM13 di-shy‐4-shy‐ANEPPS13 613 microM13 di-shy‐4-shy‐ANEPPS13

113 minute13 dye-shy‐load13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Detecting Arrhythmia Advantages for Label-Free Assessment

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

N=413 DMSO13

113 microM13 di-shy‐4-shy‐ANEPPS13

Time13 (min)13 Time13 (min)13

Detecting Arrhythmia Advantages for Label-Free Assessment

Persistent13 dye13 Serum-shy‐containing13 media13 Label-shy‐free13 assessment13 minimizes13 perturba+ons13 to13 cardiomyocytes13 13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Evoked13 field13 potenals13 in13 a13 high13 throughput13 instrument13 13

Pacing with AxIS 20

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Pacing Entraining Cultures

Cardiomyocytes13 rapidly13 entrain13 to13 electrical13 s+muli13 13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

FPD13 shortens13 as13 pacing13 rate13 increases13

Pacing Specifying Beat Period

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Changes13 in13 FPD13 occur13 over13 the13 +mescale13 of13 minuteshellip13

Pacing Practical Considerations

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

hellipbut13 are13 reliable13 across13 beats13 and13 across13 wells13

Pacing Practical Considerations

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Pacing13 enables13 accurate13 beat13 rate13 correc+on13

Control13 of13 beat13 rate13 reduces13 well-shy‐to-shy‐well13 variability13

Pacing Improving Accuracy

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Pacing13 uncovers13 reverse13 use-shy‐dependence13 of13 candidate13 compounds13

Reverse Use-Dependence

Pacing Adding Depth

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

hellip

Beat13 113 Beat13 213 Beat13 313

Beat13 413 Beat13 513 Beat13 613 Beat13 10413

Spon

tane

ous13

Pacing13

Pacing Controlling Propagation

Pacing13 establishes13 consistent13 propaga+on13 paherns13 13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

Conduc+on13 Velocity13 (mms)13

13 of13 Trials13

Paced13

Spontaneous13

Pacing Controlling Conduction

Pacing13 establishes13 consistent13 conduc+on13 measures13

2813

Questions

The13 Maestro13

2813

Questions

The13 Maestro13