CLONING: NEW FRONTIERS OF ANIMAL REPRODUCTIVE BIOTECHNOLOGY

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23 Οκτ 2013 (πριν από 4 χρόνια και 15 μέρες)

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CLONING: NEW FRONTIERS OF
ANIMAL REPRODUCTIVE
BIOTECHNOLOGY

Dr.
Gatot

Ciptadi

Lab. Of Genetics and Animal Breeding

Fac

of Animal Sci.,
UB

LSIH
-
UB

BIOTEKNOLOGI REPRODUKSI
MAMPU MENYELAMATKAN


SUMBER GENETIK

HEWAN
LANGKA

GT

Introduction

somatis

oosit

sperma


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
f
rontier of research of animal
reproductive
biotechnology


Why
?


Somatic cell (
deferentiated
) was
reprogram
ed


back to

embr
y
onal
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 y

Recipient oocytes

Donor cell

Culture of Cloned
embryos

In vitro culture

Enucleation

Introduction of donor cell

Enucleation

Passages of
somatic cells


Activation

Enbryo transfer

Production of transgenic piglets

In vitro Maturation

of oocytes

Introduction of

Extraneous gene


Xenotransplatation

Production of


human protein

Synchronization

of estrus

Surrogate mother of miniature pigs

Nuclear trasfer

Recipient oocytes

Donor cell derived
from miniature pigs

Selection of M
2

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

Enucleated

Non 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: ?