Effect of harvesting technique and presence or absence of corpus luteum on in vitro development after parthenogenetic activation of oocytes recovered from buffalo ovaries

Aim: The present study was aimed at assessing the influence of three oocyte harvesting (aspiration, puncture and slicing) methods and presence or absence of corpus luteum (CL) on recovery efficiency and subsequent in vitro maturation and parthenogenetic development of immature oocytes recovered from buffalo ovaries. Materials and Methods: Buffalo ovaries were collected from local slaughter house and oocytes were collected by using aspiration, puncture and slicing methods. Collected oocytes were matured in vitro in TCM-199 medium supplemented with 10% fetal bovine serum (FBS), 0.22 mM sodium pyruvate, 10 µg/mL of follicle-stimulating hormone, 6 IU/mL luteinizing hormone, 1 µg/mL 17-β estradiol, 100 IU/mL penicillin and 0.1 mg/mL streptomycin. In vitro matured oocytes were activated by ethanol (7%) followed by 6-DMAP (2mM) and cultured to assess the in vitro developmental capacity. Results: The average total number of oocytes recovered per ovary was significantly (P<0.05) higher in slicing (7.88±0.54) than the aspiration (2.50±0.11) and puncture (3.59±0.18) methods. The number of culture grade oocytes per ovary was significantly higher in slicing (5.40±0.29) than in puncture (2.45±0.12) and aspiration (1.94±0.11) methods. However, the percentage of culture grade oocytes was higher in the aspiration method (77.67%), compared to the puncture (68.24%) or slicing (68.51%) methods. The oocyte recovery was significantly lower (P