The Influence Of Granulose Cell On The In Vitro Maturation Of Bovine Oocytes And Development Of Early Embryos

The research of somatic cell nuclear transfer and transgentic animal has becamethe hotspot and developing direction of embryo engineering, but their effectiveness isvery low. The mainly restricting factor is the technique of IVM and IVC, the unsuitableIVP system induced poor quality and low effectiveness of IVP embryos, so it'snecessary to perfect the IVP system. The paper was undertook to study conditions ofgranulose cell monolayer co-culture with oocytes and embryos. 1 Studied the influence of granulose cell monolayer to the different grade bovine unmatured oocytes matured in vitro. The result show:The co-culture system have no influence to the maturation of grade A,B oocytes, but can obviously improve the maturation capacity of nude oocytes. 2 Probled into the influence of three Protein additions to the IVM co-culture system. Added BFF,OGS ,FBS to the co-culture system, there are no difference in cleavage rates After IVF, but blastosyst development is highest in BFF group,which obviously higher than FBS group (37.5% V S 25.2%,P<0.05), the OGS group is between BFF and FBS group,no obvious difference to both of their(P>0.05). 3 The effect of three culture medium:SOFaa,CR1aa,TCM199 were compared for the co-culture system,the rates of blastocysts were 36.08%,32.64%,11.24% respectively,the rates of hatching were 74.26%, 78.79%, 40.00% respectively.The results implied that SOFaa or CR1aa was fitter for the co-culture system than TCM199. 4 Select a protein addition for the co-culture system from FBS, OGS, BFF.The rates of blastcysts were 27.12%, 48.90%, 46.83% respectively and the rates of hatching had no obvious difference, all about 70%. The results showed that BFF or OGS was more profitable for the co-culture system than FBS 5 The effect of different co-culture periods were reseached.Zygotes were cultured in SOFaa (group1); SOFaa until Day 4, followed by SOFaa-GCM co-culture for the remainder of the culture9(group2) ; SOFaa-GCM co-culture until Day 4, followed by SOFaa for the remainder of the culture(group3) ;or SOFaa-GCM for the entire culture(group4). the results revealed that there were no obviously different in the rates of blastocysts and hatching between the group3 and group4 ,four day later's co-culture had no use for the rates of blastocysts but benefit blastocysts hatching.