Clean Meat Initiative
(Introduction)
What is cultured meat?
The technology(Questions session)
Social implications(Questions session)
“Cultured Meat” 2
“Cell culture school for kids” we organised
One of our student volunteers
“Shojinmeat Project” - Who we are 3
Collection of biohackers, science communicators, artists, student volunteers and so on, interested in cultured meat and cellular agriculture
Primary motivation in:Sustainability, culture and technology enthusiasm
Integriculture Inc. is the registered company to purchase reagents etc.
What/How we do 4
“Shojinmeat Project” develops cultured meat technology and engages in public communication.“Shojinmeat Project” is not affiliated with universities or corporations.
Low-cost culture media developed in private residence, materials from Rakuten/Amazon
Cell culture experiments in a secondhand incubator at Japanese “Indie Bio”, Leave-a-Nest Inc. lab
Self-funded, partly by SCIGRA, a science graphics & education service run by student volunteers.
Members participate in their respective expertise (experiments, gatherings, art projects etc.)
#Food Security #Food Miles
#Regulations
#Cooking
#Culture & thoughts#History#Food safety
#Life ethics#Animal welfare
#Regenerative medicine#Tissue engineering
#Bioreactor #Culture medium
#Business#LCA
#R&D
#Soc.&Econ.
Shojinmeat “Distributed Clusters” 5
#Global collab.
#Space
#Art
Shojinmeat “Distributed Clusters” 6
・Members tag themselves#R&D, #Cooking, #Ethical studies etc.
・One can even lead a cluster
・Clusters act independently, but occasionally share information
(Introduction)
What is cultured meat?
The technology(Questions session)
Social implications(Questions session)
“Cultured Meat” 7
8
“Meat”
From where?Meat is ~x40 more resource intensive
Lamb:~x50, Beef:~x40, Pork:~x20, Poultry:~x7
“Meat”←animals←feed, water, land
9
DeforestationFire-fallow cultivation Water shortage
“Meat”←animals←feed, water, land
10
Food as strategic resource and leverage
Food Security 11
“Insect Food” 12
MeatSoy etc. Dairy
MeatDairySoy etc. New alternative protein
Plants
Tofu
Algae Insects Biosynthetic Cultured
New protein source
“Meat & dairy produced in new ways”
Plant-based meat & dairy equivalent
Now
Future
“Alternative protein” and “meat alternative” 13
Tofu Plants
Algae
Insects
Biosynthetic
Quorn
Seitan
“Alternative protein” and “meat alternative” 14
Muscle cells Bioreactor
Culture medium
Processing
Cultured Meat 15
+Solution to 10000 year long ethical dilemma?
16Resources saved to 1/10~1/100
Research track record
1931 1997 2005 2013 Goldfish culture@NASA
Netherlands funds research
Prof. Mark PostDemonstration
Concept known since 19c.
17
http://www.nicovideo.jp/watch/sm30099092
18Demonstration video on Niconico
Cellular AgricultureAgricultural products by cell culture
19Medical technology, Agricultural application
¥28,000,000Cultured burger, 200g
($260,000)
The Problem 20
Culture
Cook¥3000
($28)
2121Possible. Just expensive
Blame here
22Why so expensive
Cell culture is expensive
23Culturing of cells has beenoptimized for laboratory scale
Labs, Hospitals
Brewery, Petrochemical complex
Culturing of cells has been optimised for laboratory scale
Culturing of cells becomes industrial scale
24Conditions and purposes change→Entire process needs a re-design
No 100mm dish but use 25㎥ tankNo gloves but do tank sterilisationNo pipettes but use pipelines If tanks are sterilised, no antibiotics needed
Conditions and purposes change→Entire process needs a re-design
25
・Meat production is very resource intensive・Lack of resources leads to resource war・Cultured meat is a potential solution・Possible to make, but expensive・Cell culture methodology needs redesign
What is cultured meat? (summary)26
(Introduction)
What is cultured meat?
The technology(Questions session)
Social implications(Questions session)
“Cultured Meat” 27
2.System
1.Media
3.OperationWhat needs redesign? 28
$15 $450 $410
Medium + Foetal bovine serum(FBS) + Growth factoramino acid, minerals,sugar, vitamines
Albumin, insuline, transferin
cell growth factor
Culture medium 29
= Cow blood
Standard DMEM(FBS10%) 500ml
$30,000 for 100g 500ml of culture medium produces 200mg of cells
Cost of culture media
(yen/JPY)
30
Block et al., 1996
DMEM 450ml ¥1125FBS 50ml ¥4900Non essential amino acid ¥140
HGF 40ng/ml ¥78000 (20µg) EGF 20ng/ml ¥700 (10µg)
¥84865($8000)
Culturing 100g of liver cells cost $400,000
Liver cell culture for therapy 31
SugarAmino acidsVitaminsMinerals
AlbuminBufferInsulinTransferin
Growth factorsSurvival factors
Essential medium
Foetal bovine serum
Signal compounds
Expensive… :-(“Mad cow”?Viruses?Expensive and unstable supply
Gospodarowicz D and Moran JS, 1976, Annu Rev Biochem Eagle H, 1959, Science
Expensive for what’s actually in
=
Bottlenecks in culture media 32
Our research direction 33
・Biohack.Do things do-able without privilege
・Start with something radically low-cost Such materials become animal-free by economics
・No jumping to making meatFind a cheap way to do it first.
Yeast extract as FBS alternative? 34
Commercially available:“Yeast extract for animal cell culture”
(Sheffield Bioscience Ltd.)
Dried yeast (dog food)
Phosphate buffer & saltPapain, 65℃, stir, ~24hto “digest” yeast
Filtered before use in cell culture
pH7.4No bacterial contaminationPreparation cost: 10¢/L
Myoblast cell response 35
The trial solution showed signs of L6 growth and multiplication. ⇒”Yeast extract (dog food) is a potential FBS alternative.”
NO
FB
SC
ontr
ol
(with
FB
S)Mouse L6 Myoblast cell density (qualitative)Red ~ Yellow ~ Blue(zero)
No DMEM
w/ DMEM
No DMEM
w/ DMEM
Conc. yeast extract
Dil. yeast extract
HeLa cells in yeast extract medium 36
HeLa cells in sYE (yeast extract) multiplied at 80% of that in FBSHeLa cells in sYE medium grew in diameter more than in FBS
Cel
l cou
nt (x
10e5
)(HeLa = Human cervical cancer cell line)
Day0 Day7
FBS 10%
sYE 10%
sYE 10% +FBS 10%
293T cells in yeast extract medium 37
sYE is capable of culturing 293T cells, which have relatively strict requirement for FBS to thrive
Cel
l cou
nt (x
10e5
)(293T = Human Embryonic Kidney cells)
Day0 Day7
FBS 10%
sYE 10%
sYE 10%+FBS 10%
10% FBS
10% sYE
10% FBS10% sYE
Cellular self-organisation in sYE 38
96-well suspension culture plate
Mouse myoblasts were multiplied to 1.0E5 in standard FBS medium Cells were placed under suspension culture condition in sYE medium for 4 days⇒Self-organised into a 3-dimensional aggregate of 0.3~0.5mm diameter
The limits 39
Species Cell identity sYE conc. Result
Human HeLa Human cervical cancer 10% Successful (but for how long?)
Human Human Embryonic Kidney 293T 10% Successful (same as above)
Mouse Germ cell 10% Unsuccessful
Mouse Ovarian somatic cell, primary 10% Successful (but for how long?)
Mouse Hepatocyte, primary 10% Successful (same as above)
Mouse Fetal cell, primary 10% Successful (same as above)
Mouse Myoblast, primary 10% Initially successful, but cell division halts after ~3gen.
Mouse Embryonic stem cells 10% Unsuccessful
Yeast extract do have limits.Unsuitable for undifferentiated cells.
2.System
1.Media
3.OperationWhat needs redesign?
Breaking the limits 41
How do body fluids acquire albumin, growth factors etc.?⇒Intercellular interactions
Ohlsson C et al., 2009, Endocr RevFrancis GL, 2010, Cytotechnology
HGFEGF
TGFβAlbumin
Adrenalin
Coculture system (proto-prototype)
PCT/JP2016/067599Japan pat. 2016-568716
Conditioning of culture media by coculture 42
Control (0%) 10% conditioned medium
25% conditioned medium
50% conditioned medium
Cou
nt o
f cel
ls o
f all
size
s,
rela
tive
to th
e co
ntro
l gr
oup
Secondhand medium is effective!
mouse placental cells
dishes with Day-12 foetal liver cells in FBS 10% medium
7 Days
Transfer culture medium
Low-cost liver cell culture 43
DMEM 450ml ¥1125Non essential amino acid ¥140FBS 50ml ¥4900HGF 40ng/ml ¥78000 (20µg)EGF 20ng/ml ¥700 (10µg)
¥84865(€750~)
Liver cell aggregate on collagen scaffoldCultured by coculture in sYE medium
DMEM 450ml ¥1125sYE 50ml ¥182
¥1307 (€12)
The full potential of “sYE in coculture” 44
DMEM 450ml ¥1125sYE 50ml ¥182
¥1307 (€12)
”DMEM” 450ml ¥10sYE 50ml ¥1
¥11 (€0.10)
SugarAmino acidsVitaminsMinerals
Essential medium =
from algae?
MediumGO 45
Huge variety on streets to choose from!
The cost of media for 100g of cells 46
Current method cultures only 1 layer of cells on the bottom.
Improvements in culture efficiency 47
Scaling as it is... 48
How cultured burger was made in 2013⇒$260,000
Suspension culture Cellular scaffolds
More efficient methods 49
Either way is called “3D culture”, not bound on 2D dish
FIRST Project by Prof. OkanoSystem integration for commerci-alisation of regenerative medicinehttps://www.youtube.com/watch?v=g7lyTsxbt2U
Suspension (3D) culture uses less medium
Culture media
Culture media
27 Litres
9 Litres
What it takes to grow 1B iPS cells
Efficiency by suspension culture 50
Cellular scaffold also achieves the advantage of 3D culture
Sponge collagen scaffolds Liver cells on scaffolds
3D culture by cellular scaffolds 51
Edible scaffolds i.e. collagen, chitosan, chitin, arginate, cellu- lose, polysaccharides
Simulate fibre and meat texture
Moulds shape in mm or bigger scales
Other benefits of scaffolds 52
Integration of all elements into one scalable system
~0.1g scale ~10g scale ~100kg scale future pilot plant
All in one system: sYE, scaffold, suspension 53
PCT/JP2016/067599 jp-pat file# 2016-568716
All in one system: sYE, scaffold, suspension
technologically demanding“Is it tasty?” 55
“Ground meat” “Meat sheets” “Steak”At least it’s meat Juicy tastes Meat textureMade by muscle +Fat cells +Blood vessels,+fat cells Regen. MedicineProven ※though expensive Scaffolds
2.System
1.Media
3.OperationWhat needs redesign? 56
Not at all big enough for food
57Current biggest cell culture
Plant engineering 58
How is temperature controlled?
Mixing method?
Pipeline diameter & flow rate?
Sterilization method & frequency?
How are filters cleaned?
Plant engineering - what exactly? Speculative fish meat culture plant
・Resource requirement from ‘cradle to the grave’
・LCA is necessary for plant engineering
Life cycle assessment (LCA)
“Meat brewery” 61
Farm high-rise 62
Mars Colony 63
Orbital Zero-G Farm 64
(Introduction)
What is cultured meat?
The technology(Questions session)
Social implications(Questions session)
“Cultured Meat” 65
Would vegetarians eat?Is it Halal?Goes with Buddhist “nonviolence”?Animal welfare?Consumer acceptance?At the end, is it tasty?
Social and cultural implications 66
A. Animals, religion B. Food security C. Food safety
D. Env. footprint
EU/US: A~D > C > BE.Asia: B~C > A > D
Regional differences in agenda 67
MENA & India: i.e. Halal/Kosher 68
・Yehuda Shurpin, a Jewish cleric: “If the original cell is “Kosher”, so would be the cultured meat, but let’s wait until technologies mature.”・Abdul Qahir Qamar of Jeddha Islamic Academy: “food from animal secretes - isn’t it just yoghurt?”・Holy Cow for for-profit operations? NO, killing or welfare isn’t the issue・Pork is “unclean” because of its lifestyle and habitat? ….or the DNA?
Utilitarian & Animal welfareBuddhist “nonviolence” commandmentGeneral consumer acceptance?“New Ethics” controversy in 2040
Selected topics 69
Personal beliefs than religions
・Vegetarians・Pescetarians・Vegans are the most common
Western vegetarianism 70
“Max. happiness for the max. number of sentient beings”※Not “ends justify means”, but an guide to arguments over what’s ethical
“Only 1 death is better than 5”...?May be so in very short term, but...
What if the person is your brother? A society where abandoning of family is justified, would it be a good one?
or in another example,If “killing for public good” becomes the norm, no one is there to stop dictators - would it really do “max. happiness for max. number of people? ”Long-term total happiness must be discussed”
Classic “Runaway trolley problem”As it is, 5 will die, but if switch, 1 dies. Would you switch?
Practical ethics: “Utilitarianism” 71
“Animal welfare” deduced from utilitarianism 72
All interests are equal: “1 person, 1 vote”No being’s happiness is more important than another
◆Are animals capable of experiencing happiness?◆Should an animal count as one sentient being?
In “animal welfare”, speciesism is dismissed & animals count as beings that can experience happiness
⇒From a utilitarian point-of-view, “making a sentient being suffer is unethical”
Acknowledges health and environment, but mainly animal welfare and ethical
“Animal welfare” based on utilitarianism
Reasons for vegetarianism 73
Announced in 2008 ⇒ taken down on 2014: technology wasn’t ready
$1M Award by PETA 74
Interests inAnimals⇒↓Veggie
YES NO
YES Vegans, mainstream Western vegetarians
Opinions divided
For-health veggies, Pescetarians
“Not for me”...?But beef meat fish fat “healthy meat” maybe?
NO Animal welfare organisationsPet owners, animal lovers
All-out YES
General consumers
What is it? GMO? Zombie meat?
Opinions and positions 75
”Religion”←Not utilitarianism or other ethics
“Ethical” aspects of cultured meat don’t necessary translate into East Asian religious (i.e. Buddhist) importance.
East Asian vegetarianism 76
Cuisine for for zen practitioners. Its preparation is also part of zen practice.
・Common name for “Buddhist cuisine”・Uses only local produces・Donated food - use without waste・No use of meat and several specific vegetables.※If meat is donated, it is cooked and consumed.
Cultured meat doesn’t equal “shojin ryori” solely on the grounds of “because it hasn’t killed” - it must carry more religious meanings.
“Shojin ryori” Buddhist cuisine
Holy text↓
Rules↓
Daily life
Religious rules in Abrahamic religions
RulesMiddle way
Daily life
Holy texts
・Middle way > Rules・Making religious rules as something “absolute” goes against the middle way.
・Nonviolence is one of the rules
・Monks eat meat, if donated.
・Cultured meat would be seen as a kind of “tofu (fake meat)” Use it as a reference
in making decisions
Mentions
Religious rules in Eastern religions
“The Middle Way”
Before 7th century:Eating meat was common. People just had to eat whatever was in hand.
675c. Imperial decree of “No Killing (of animals)”To direct labour force to rice production and put a stop to local animal-sacrifice rituals & reinforce imperial authority※Newly arrived Buddhism was used as justification
Meat avoidance continues till 19th c. and commoners only started eating meat around 1900 c.
Meat in historical Japan
Totals half billion? Region-specific
Found more in upper caste, which even funds cultured meat research
Hinduism doesn’t explicitly forbid, but adherents choose to avoid meat.
Current Indian PM is one.
Vegetarianism in India 80
Guardian, 2014/09/11
(2014/08/05)
Public opinion is fluid due to lack of information on the subject.
Very fluid
General consumer acceptance
NO YES
(Transient) issues upon R&D:・FBS production is not “cruelty-free”
“Unavoidable” issue:・Extraction of cells
Likely to be solved in the futureMay pose an issue during R&DCan bovine foetus feel pain?
It may still inflict some pain.Will animals still be chained?Genetic selection of animals for the sake of “tasty” - is it eugenics?
Ethical issues due to technological immaturity
Then what if on 2040, cultured meat takes up 30% market share and more people stop consuming live animals for meat?・Uncontrollable ”hate campaign” against traditional farmers?・Trade ban of “real” meat by WTO on animal rights grounds?
Why are animal experimentation, Japanese whaling and Chinese cat/ dog consumption is problematised far more than factory farming? ⇒Because they are “remote things” for the protesters.
Possibility of “New Ethics”
・Can countries with meat culture technology blame others for animal abuse?
・”Patent infringement to save animals in countries that don’t have the technology”
Case study: Generic HIV drug lawsuit:
An Indian pharmaceutical company allegedly infringed retroviral drug patent to manufacture generic HIV drugs, because the original drugs by Western pharmaceutical companies were too expensive for people in poor African countries.
After high-profile court-martials, the Indian company won the case on humanitarian basis.
Patent war under “New Ethics”
Global meat$1.5T
Global beef$0.7T
Japan$50B
Toward “price parity”
Startups
Funds
NPOs
Key start-ups and NPO’s
Members participate in their respective expertise (experiments, gatherings, art projects etc.)
#Food Security #Food Miles
#Regulations #Cooking
#Culinary culture#History#Food safety
#Life ethics
#Animal welfare#Regenerative medicine#Tissue engineering
#Bioreactor #Culture medium
#Business
#LCA
#R&D#meat culinary culture#Soc.&Econ.
Shojinmeat “Distributed Clusters” 86
Acknowledgements 87
Lab space, various technical & management advices
(Ms. Murata’s private residence)
Members & Volunteers
Dr.Komatsu Research Office
“Science Agora” event organization
“Homebase”meeting & event space
Support for our “Cultured meat fanzine” distribution
Niconico science community
Miyo-sanFrom ”Myosin”
age.20・164cm
Chemical engineering student intern at Mars Huygens Crater cellular agriculture plant
Aco-chanFrom ”Actin”
age.13・149cm
Helps elder sister Miyo as part of extracurricular activity offered by Mars Colonists Middle School.
Miyo & Aco
Miyo-san (Chibi)
Aco-chan (Chibi)
Contacts 93
http://[email protected]
http://[email protected]
http://[email protected]
Scientific VR, 3D printing and rendering service -funds Shojinmeat Project
http://lne.stSeed accelerator