P-34: Effect of Including Growth Factors andAntioxidants in Maturation Medium Used forIn vitro Culture of Buffalo Oocytes RecoveredIn vivo

Background: This study examined the effect of including one of two growth factors (100 ng/ml IGF-1 or 20 ng/ ml EGF) in combination with one of two antioxidants (50 μM cysteamine or 50 μM beta-mercaptoethanol) in invitro maturation of buffalo oocytes, followed by fertilization and subsequent development of embryos. Materials and Methods: The oocytes were recovered by in vivo ovum pick-up technique from six Murrah buffalo heifers twice a week over a period of 16 weeks. Immediately after ovum pick-up, oocytes recovered from six donors were allocated randomly to five different maturation treatments. The control treatment was the basic maturation medium (MM; TCM-199 supplemented with 10% FBS, 10 IU/ml LH, 0.5 μg/ml FSH, 1 μg/ml estradiol- 17beta and 50 μg/ml gentamicin). The other four treatmentsconsisted of the control maturation medium (MM) plus one combination of a growth factor and an antioxidant viz. IGF-1 + cysteamine; IGF-1 + beta-ME; EGF + cysteamine or EGF + beta-ME. The total number of oocytes assigned to each maturation treatment ranged from 31 to 66. After maturation in different maturation medium, media used for in vitro fertilization was same for all groups i.e. mSOF along with some supplements. Following in vitro fertilization, the presumptive zygotes were stripped of the cumulus cells and were cultured for subsequent development over cumulus cells monolayer along with active motile bovine oviductal epithelial cells in supplemented mSOF. Data were analysed using Chisquare test.Results: The maturation rate observed for the growth factor plus antioxidant treatments was similar to that for the control (90.4%). The highest cleavage rate recorded in the IGF-1 + cysteamine treatment (71.9%) which was significantly higher (p