| The effects of follicular fluid, sperm-oocyte co-culture time, volume of sperm-oocyte co-culture fluid, layer of cuculum and season were investigated. And the effects of freezing method, freezing carrier, anti-freeze and time of blastocyst were studied in present experiment.Preliminary study on optimization for system of bovine oocytes IVF: (1) The cleavage rate of fertilization for 10 h is better than that in 7 h, there was significant difference between fertilization for 10 h to 7 h (P<0.05), and there was no significant difference between fertilization for 10 h to 7 h(P> 0.05) in hatching rate. (2)the cleavage rate of mature fluid added SFF and BFF were 75.41﹪and 63.01﹪respectively, there was significant difference between BFF to SFF (P< 0.05).Study on vitrification of IVF embryo in boving: (1) There were no significant difference in Full shape rate between EG to EG + DMSO (P> 0.05). There was significant difference in hatching rate between EG to control (P< 0.05). There were no significant difference in hatching rate between EG to EG + DMSO (P> 0.05). (2) the hatching of earlier blastcyst which were vitrification in OPS and procedure freezing were not better than that of control (30.23%, 35.56% and 39.34%), but there were no significant difference (P> 0.05). It showed that the effect of earlier blastcyst which were vitrification in OPS and procedure freezing was all better. (3) There was extremely significant difference in Full shape rate between GMS to 0.25ml straw (P< 0.01). There was significant difference in Full shape rate between OPS to 0.25ml straw (P< 0.05). But there were no significant difference between OPS to GMS (P> 0.05). It showed that there was not good in 0.25ml straw, OPS and GMS were all good, and it is possible OPS instead of GMS. (4) There were no significant difference in Full shape rate between morula to blastcyst (P> 0.05). There was significant difference in hatching rate between morula to control (P< 0.05). There was significant difference in hatching rate between morula to blastcyst (P< 0.05). (5)There were no significant difference in Full shape rate between morula to early blastcyst (P> 0.05), There was significant difference in survival rate between pre-thawed morula to post-thaw morula (P< 0.05).Preliminary study on optimization for system of sheep oocytes IVF: (1) the cleavage rate of mature fluid added SFF and BFF were 65.23﹪and 60.43﹪respectively, there was no significant difference between SFF to BFF (P> 0.05). And that Blastocyst rate were 26.16﹪和17.11﹪respectively, there was significant difference (P<0.05). (2) The cleavage rate of fertilization for 22 h , 18 h and 12 h were 64.85﹪, 54.39﹪and 48.72% respectively, there was significant difference between fertilization for 22 h to 18 h (P<0.05), and there was extremely significant difference between fertilization for 22 h to 12 h(P< 0.01). (3)The cleavage rate and blastocyst rate of 100ul volume of sperm-oocyte co-culture fluid were better than that of 70ul and 200ul, but there were no significant difference (P> 0.05). (4) The cleavage rate and blastocyst rate of 1~3 and 3~5 Layer of cuculum were better than that of 0 layer, there was significant difference between 1~3 layer to 0 layer (P<0.05), but there was no significant difference between 3~5 layer to 0 layer (P> 0.05). (5) Cleavage rate and blastocyst rate in autumn were higher than that in summer, and there was significant difference between autumn to summer (P<0.05), but there was no significant difference between autumn to spring (P> 0.05).In conclusion, the system of sheep IVF and vitrification of IVF embryo in boving were established in my Laboratory. |
