Effects Of Granulosa Cells,follicular Fluid On In Vitro Maturation And In Vitro Fertilization Of Goat Follicular Oocytes

The present study was conducted to investigate the effects of granulosa cells\follicular fluid on in vitro maturation and in vitro fertilization of goat follicular oocytes.The main results are as follows:The IVM rates of cumulus-oocyte-complexes (COC5) .. denuded oocytes (Do) and naked oocytes (No) were 67.68%.. 45.3%.. 7.3%, respectively. There was no significant difference among all groups. After the matured oocytes were inseminated in vitro , The 2-cell cleavage rates were 14.1%.. 2.68%.. 0. respectively. The 2-cell cleavage rate of COC5 was significant higher than that of Do and No.The cleavage rates of maturated oocytes with expanded cumulus cells or with complete cumulus denudation were 14.1% and 14.32%, There was no significant difference between two groups.After cultured for 24h in maturation medium with 0.1 Sx I O6viable cells/mi(I).. 1.5x 106 viable cells/ml(ll) .. lSxlO6viable cells/ml(Lll).. 150X 106 viable cells/ml(JV)of cumulus granulosa cells, the maturation rates of COC5 were 60.0%.. 64.37% 50.15% .. 30.4%,respectively.Group TILT and IV were significantly lower than control group (65.4 l%).After IVM, all maturated oocytes from each group were inseminated ,the 2-cell cleavage were 14.2%.. 14.6%.. 12.47%.. 12.56%.. 13.4%(control group) ,respectively..There was no significant difference among all groups. The effect of follicle granulosa cells on IVM and IVF had also been examed and we had the same results as cumulus granulosa cells. There was no significant difference between them.After cultured in monolayer medium of cumulus granulose cells or in inconditioned medium of cumulus granulosa cells for 24h. the maturation rates of oocytes in monolayer medium was significantly higher than that of conditioned3medium(66. 10% vs 44.2%).There was no significant difference between the monolayer group and the control group(66.10% v~ 66.28%).The effect of goat follicle fluid (GFF) on the DIM and IVF of COC5 were examed. Liberated COC5 were cultured either alone in GFF or in M199. After 24b~the maturation rate of oocytes cultured in GFF was significantly lower than those cultured in M199.When the COC5 were transferred to medium without GFF and incubated for an additional 24h,62.34% of COC5 were matured and there was no significant difference in maturation rates between conditioned and control groups.After WF,2-cell cleavage rates were 14.5% and 12.490/o,respectively.there was no significant difference between two groups.GFF was collected at different stages of the estrous cycle from small follicle (2-3mm)~ medium follicle(3-Smxn) and large follicle (5-6mm),then all oocytes were incubated in each groups. After cultured for 24h,the FF from any follicle size at follicle phase had the least DIM inhibition activity. However to the same follicle size, FF from early luteal phase(group I) and late luteal phase(group IV) had significant lower DIM inhibition activity than luteal phase(group 111 ).There was no significant difference between group II and IV.Except at follicle phase,the DIM inhibition activity of (3FF at the same stage did not differ with different follicle size. At follicle phase ,the DIM inhibition activity of GFF significantly decline during the course of follicular development in small medium and large follicle.Added o%~ 5%.~. l0%.~ 15% (3FF to the maturation medium ,the maturation rates were 66.4% 64.7%~ 64.3%.. 60.6%,respectively after incubated for 24h,and there was no significant difference among all groups. After inseminated , 2-cell cleavage rates were 13.57% .. 14.0% 13.4%13 .48%,respectively. There was also no significant difference among all groups.