VPB 321: Animal Biotechnology (2+1)

IN VITRO CULTURE AND MATURATION OF OOCYTES

• Oocytes for in vitro fertilization are obtained from the oviducts, follicles and surface of ovary after ovulation either from live animals by flushing some medium or from slaughtered animals. The immature oocyte is collected from the ovary and cultured in vitro to induce maturation.
• Cow ovaries are collected from the slaughter house and maintained in phosphate buffered saline at 30-32°C for transportation to the laboratory. Following aspiration of follicles, the oocytes are then observed under a dissecting microscope.
• The oocytes are graded according to their morphology, washed twice with HEPES buffer and once with BMOC-3 (Branchat and Olivhant medium for oocyte collection). After washing, 5-10 oocytes are transferred to 0.2 ml of BMOC-3 drops under paraffin oil at 37°C with 5% CO₂ in 95% air for 12-32 hrs.

Oocyte maturation in vitro

• Large numbers of oocytes are allowed to mature under in vitro conditions.