CLONING: NEW FRONTIERS OF
ANIMAL REPRODUCTIVE
BIOTECHNOLOGY
Dr. Gatot Ciptadi
Lab. Of Genetics and Animal Breeding
Fac of Animal Sci., UB
LSIH-UB
Introduction
somatis
oosit
sperma
BIOTEKNOLOGI REPRODUKSI
MAMPU MENYELAMATKAN
SUMBER GENETIK HEWAN LANGKA
GT
Basic technique for nuclear transfer
(cloning)
1. Cryopreservation (Freezing) Seeding, Stepwise, Direct, Vitrification
2. Embryo transfer (ET)
Superovulation
Collection of embryos (Flushing)
Evaluation of embryos
Synchronization of estrus
3. In vitro fertilization (IVF) Collection of oocytes from ovaries
In vitro maturation
In vitro fertilization
In vitro culture
4. 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.
Bioteknologi Reproduksi: Sumbangan KLONING
Pada Ilmu Pengetahuan
GT
GT
Summary
Recipient oocytes
Invitro maturation
Enucleation
Donor cell
Nuclear transfer
In vitro culture
Activation
Culture of Cloned
embryos
Surrogate mother pig
ET
Nuclear trasfer
Recipient oocytes
Donor cell derived
from miniature pigs
Activation
In vitro Maturation
of oocytes
Passages of
somatic cells
Surrogate mother of miniature pigs
Synchronization
of estrus
Selection of M2
Introduction of
Extraneous gene
Enbryo transfer
Production of transgenic piglets
Enucleation
Xenotransplatation
Introduction of donor cell
Production of
human protein
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
TECHNICAL ASPECT: TN
Non enucleated
Enucleated
IDNI
Enucleation
Seleksi Sel Donor
Ukuran sel
1
2
3
4
A
1. Besar,
B
2. Sedang,
Sel Hidup… / Mati…
3. Kecil: 22.16 %
4.Abnormal
Tanpa Aktivasi
Aktivasi tunggal
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: ?