CLONING: NEW FRONTIERS OF ANIMAL REPRODUCTIVE

BIOTECHNOLOGY

Dr. Gatot CiptadiLab. Of Genetics and Animal Breeding

Fac of Animal Sci., UBLSIH-UB

BIOTEKNOLOGI REPRODUKSI MAMPU MENYELAMATKAN

SUMBER GENETIK HEWAN LANGKAGT

Introduction

somatis

oosit

sperma

Basic technique for nuclear transfer (cloning)1. Cryopreservation (Freezing) Seeding, Stepwise, Direct, Vitrification2. Embryo transfer (ET) Superovulation Collection of embryos (Flushing) Evaluation of embryos Synchronization of estrus3. In vitro fertilization (IVF) Collection of oocytes from ovaries In vitro maturation In vitro fertilization In vitro culture4. Micromanipulation Sperminjection (ICSI; IVM, Activation, IVC) Clone (Nuclear transfer) IVM, IVF, IVC Enucleation, Nuclear injection Fusion, Activation Gene transfer ( vector, Electropolation) Gene transfer (Injection into zygotes)

Cloning Techniques

Desection of morula and blastocyst (Embryo) Deaggregation (Isolation) of blastomeres (Embryo) Clone Micromanipulation Nuclear transfer Micromanipulation +Gene transfer

1. Desection of morula and blastocyst (Embryo)

2. Deaggregation (Isolation) of blastomeres (Embryo)

• DREAMS SOMETIMES COME TRUE !

Bioteghnology has the possibility to be revived a scare and an extinct animal (Film Jurasic Park)

• The birth of the clone animals is great influencing the frontier of research of animal reproductive biotechnology

• Why ?• Somatic cell (deferentiated) was reprogramed

back to embryonal stage and resulted in normal offsprings.

GT

Bioteknologi Reproduksi: Sumbangan KLONING Pada Ilmu Pengetahuan

GT

Nuclear transfer Activation

ET

Invitro maturation

Surrogate mother pig

S u m m a r yRecipient oocytes

Donor cellCulture of Cloned embryos

In vitro cultureEnucleation

Introduction of donor cell

Enucleation

Passages of somatic cells

Activation

Enbryo transfer

Production of transgenic piglets

In vitro Maturationof oocytes

Introduction of Extraneous gene

XenotransplatationProduction of human protein

Synchronization of estrus

Surrogate mother of miniature pigs

Nuclear trasfer

Recipient oocytes Donor cell derived from miniature pigs

Selection of M2

Nuclear transfer procedure to produce reconstructed embryo: inject donor cell in the perivittelin space of enuclated oocyte • the performed fusion cell is performed• intracytoplasmic direct nuclear injection (IDNI)• activated artificially

EnucleatedNon enucleated

IDNI

Enucleation

TECHNICAL ASPECT: TN

    

  1

2

3

4

A B 

Sel Hidup… / Mati…

1. Besar,

2. Sedang,

3. Kecil: 22.16 %

4.Abnormal

S e l e k s i S e l D o n o r

         

Tanpa Aktivasi Aktivasi tunggal

Ukuran sel

DEVELOPMENT OF NEW TECH. IN REPROD ANIMAL

1 1997 Birth of dolly (1 st animal cloning using somatic cells)

2 1998 Birth of cows: Charlie/George: ( serum albumin)Specific protein for human blood aglutination

3 2000 Pig cloning (transgenic) for organ transplantation (human),

4. 2001 Inter species nuclear trasnfer (Yak to Bos Taurus)

advantages: non conventional product of livestock:- Genetic quality improvement- Biopharmacies- Organ Transplantation (Biomedic industry)- Genetic concervation of endangered animals

Genetic Consideration and Constraints of Gen Manipulation???:

No. Persilangan Fertilitas Keunggulan F1

Jantan Betina

1. BantengX Zebu - + Laju pertumbuhan

2. Keledai Xkuda - + Kerja + ketahanan

3. Sapi x Yak - - BB/Power/Susu

4. Bos taurusxB. Indicus + + Daging

5. Entog X Itik ? ? Daging /pertb.

Catatan:• Bukan penghasil bibit• Secara teknis memungkinkan, tetapi• Tujuan utama bukan genetik .

Have a nice dream………

Dr. Tanaka H.

CONCLUSSION:Result from IVM, IVF, IVG of germ cell are promising What Further research: ?