General Procedure Collection of immature oocytes The maturation of immature oocytes(IVM) The fertilization of mature oocytes(IVF) The culture of embryo(IVC)

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Presentation transcript:

General Procedure Collection of immature oocytes The maturation of immature oocytes(IVM) The fertilization of mature oocytes(IVF) The culture of embryo(IVC)

Oocyte collection Immature oocyte in form of cumulus-oocyte complex What is the source of collection? Directly from ovaries obtained from abattoir From donor animal Either by laparotomy Aspiration using syringe and needle Using transvaginal ovum pick up techniques

Transvaginal ovum Pick up(OPU)

Oocyte collection by slashing ovary

Oocyte maturation Placement in maturation medium Maturation medium(pyruvate, glucose, glutamine, serum), hormones and gonadotrophins,(estrogen, LH, FSH) and antiboitics(pencillin,streptomycin, or gentamycin) Cow- it will take 21-24 hrs

In vitro fertilization COC washing Fertilization Alternative to IVF-ICSI By injecting 1 selected sperm cells into 1 mature oocyte Embryo culture- 7days Grade -1 and grade-2 blastocyst are usually transferred to recipients

In vitro criteria for quality of embryo Excellent: spherical, symmetrical with cells of uniform size Good: few extruded blastomeres, irregular shape Fair: Extruded blastomeres, degenerated cells Poor: Numerous extruded blastomeres

Evaluation of embryo development 2-16 cell stage Morula: Embryo with 32 cell stage Compact morula: Compaction of balstomeres Early blastocyst: Formation of blastocoel has just started Blastocyst: well defined blastocoel start differentiation Expanded blastocyst: Expanded with zona thinning Hatched blastocyst: partly or completed come out from rupture zona

Cell stage and estimated age in days Morula- 5 days Compact morula-6 days Early blastocyst- 7 days Balstocyst-7 days Expanded blastocyst- 8 days Hatched blastocyst- 9 days

Advantages/comparison with ETT Animal can be harvested at very young age Animal can be harvested every 20 days. It reduces the generation interval for animals with specific desirable trait No need to superovulate No need to synchronization Less no. of straws used.

Disadvantages It is more costly. Need more lab equipment. Need more laboratory skills Trained staff