| In vitro fertilization is an earlier research technique. It has been developed and widely used in production. Micro injection technology has been widely studied in experimental animals such as frogs and mice, and the research results of the cattle and sheep were significant.In this study, we chose cattle as the object of study, further research in vitro fertilization technology and micro injection in order to improve the efficiency of in vitro fertilization, explore the optimal injection pressure of the micro injection,and then to provide a theoretical basis for breeding and improving of cattle. The results are as follows:(1) Through comparison of maturation rates of different mature culture mediums, we make sure that optimal culture system of oocytes in vitro maturation is 80%M199+ 20% FBS+FSH(1.2 IU/mL)+LH(0.12 IU/mL)+E2(5 mg/ml)+100 IU/mL,maturation rate was about 80%;(2) By comparing the fertilization rates of different semen and by semen, and development rate of the developed in different ways, we finally get the best fertilization and development system, that is, to take the epididymal sperm, the concentration of sperm was adjusted to 1 x 105/ml, use the finished semen to fertilizing, the fertilization rate reached about 70%, the development system use SOFaa, and on the third day the culture medium was changed with the new which was supplemented with 10% FBS, blastocyst rate is close to 20%,however the rate of co-culture efficiency is O;(3) Choose enzyme-free TE buffer as injectant. we compared fertilized eggs development rates between the control group and different injection pressure groups. The result shows that compared with the control group, with 40hpa injection pressure fertilized egg cleavage rate and blastocyst rate difference was not significant (P>0.05), but 30hpa,50hpa,70hpa and 100hpa injection group cleavage rates had significant difference (P<0.05), under the pressure of 70hpa and 100hpa blastocyst rates had extremely significant difference (P<0.01). |
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