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CLONING: NEW FRONTIERS OF ANIMAL REPRODUCTIVE BIOTECHNOLOGY

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.

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CLONING: NEW FRONTIERS OF ANIMAL REPRODUCTIVE BIOTECHNOLOGY

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  1. CLONING: NEW FRONTIERS OF ANIMAL REPRODUCTIVE BIOTECHNOLOGY Dr. GatotCiptadi Lab. Of Genetics and Animal Breeding Fac of Animal Sci., UB LSIH-UB

  2. Introduction somatis oosit sperma BIOTEKNOLOGI REPRODUKSI MAMPU MENYELAMATKAN SUMBER GENETIK HEWAN LANGKA GT

  3. 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. MicromanipulationSperminjection (ICSI; IVM, Activation, IVC) Clone (Nuclear transfer) IVM, IVF, IVCEnucleation, Nuclear injection Fusion, Activation Gene transfer ( vector, Electropolation) Gene transfer (Injection into zygotes)

  4. 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)

  5. 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.

  6. BioteknologiReproduksi: SumbanganKLONING PadaIlmuPengetahuan GT GT

  7. S u m m a r y Recipient oocytes Invitro maturation Enucleation In vitro culture Donor cell Nuclear transfer Activation Culture of Cloned embryos Surrogate mother pig ET

  8. Nuclear trasfer Donor cell derived from miniature pigs Recipient oocytes Activation In vitro Maturation of oocytes Passages of somatic cells Surrogate mother of miniature pigs Synchronization of estrus Enbryo transfer Introduction of Extraneous gene Selection of M2 Production of transgenic piglets Enucleation Production of human protein Xenotransplatation Introduction of donor cell

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

  10. TECHNICAL ASPECT: TN Non enucleated Enucleated IDNI Enucleation

  11. S e l e k s i S e l D o n o r Ukuran sel 1 2 3 4 B A 1. Besar, 2. Sedang, 3. Kecil: 22.16 % 4.Abnormal Sel Hidup… / Mati… Aktivasi tunggal Tanpa Aktivasi

  12. DEVELOPMENT OF NEW TECH. IN REPROD ANIMAL • advantages: non conventional product of livestock: • Genetic quality improvement • Biopharmacies • Organ Transplantation (Biomedic industry) • Genetic concervation of endangered animals

  13. Genetic Consideration and Constraints of Gen Manipulation???: • Catatan: • Bukan penghasil bibit • Secara teknis memungkinkan, tetapi • Tujuan utama bukan genetik .

  14. Have a nice dream……… Dr. Tanaka H. CONCLUSSION: Result from IVM, IVF, IVG of germ cell are promising What Further research: ?

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