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

4. 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

5. Transvaginal ovum Pick up(OPU)

6. Oocyte collection by slashing ovary

8. 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 hrs

9. 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

10. 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

11. 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

12. 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

13. 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.

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