Study On Four Key Technology In Vitro Fertilization Of Bovine

The main objective of this study was to optimize IVP system of bovine embryoThe ability of in vitro maturation, fertilization of bovine oocytes derived from interior ovary (IO oocytes) was studied. The result showed that IO oocytes co-cultured with oocytes from the follicles which diameters were 2-8mm could obviously improve rate of cleavage and development of 6-8cells (P<0.05); Different concentrations (10%, 20%, 30%) of BFF were supplemented to the maturation medium, addition BFF had similar rate of cleavage and development of 6-8cells compared to no BFF control group (P>0.05). BFF could support in vitro maturation of IO oocytes and subsequent development capacity. Adding of 10% BFF could obtain the best purpose. Removal of cumulus cell prior to insemination significantly reduced the rate of cleavage (P<0.05).Sorted sperm was disposed by Percoll and swim-up methods. The result showed that the rate of cleavage and blastocyst between Percoll method and swimming-up method had no significant difference (P>0.05). The rate of cleavage and blastocys had no striking difference between sorted sperm and unsorted sperm.The effects of different concentration of taurin on in vitro development of bovine embryos were studied. The result showed that addition taurin could enhance development of bovine embryos, but addition taurin group had no significant difference than no taurin group. Addition 7mMtaurin is best choice. The effects of different concentration of amino acids on in vitro development of bovine embryos were observed. The result showed aptitudinal proportion of amino acids could improve development of bovine embryos. 3×EAA+1×NEAA was optimum choice. The effects of both TCM199 medium and mSOF medium on in vitro development of bovine embryos were studied. The result revealed that both TCM199 medium and mSOF medium could promote development of bovine embryos. The rate of cleavage and blastocyst between TCM199 medium and mSOF medium had no significant difference (P>0.05). To development of bovine embryos, mSOF medium may substitute TCM199 medium.Twain different culture medium was used to study the in vitro culture of the preantral follicles from bovine ovaries. The results showed both mediums may promote development of preantral follicles, but the rates of antrum formation and viability had no obviously difference (P>0.05). Effects of the number of follicles in each well (one follicle in each well. Two follicles in each well, three follicles in each well) on in vitro development of preantral follicle were studied. The results revealed the rates of antrum formation and viability had no significantly difference (P>0.05).