牛体外受精卵的二步法培养体系的研究

以CR1aa为基础培养液,采用二步法对牛体外受精卵进行体外培养,完善牛体外受精卵的培养体系.实验一:对照组连续7 d均为CR1aa 50 mL/L FBS培养,处理组前3 d为CR1aa 3 mg/mLBSA培养,后4 d换为CR1aa 50 mL/L FBS.处理组的囊胚率显著高于对照组,但卵裂率和囊胚孵化率无显著差异.实验二:对照组连续7 d均为CR1aa 50 mL/L FBS培养,处理组前3 d为CR1aa培养,后4 d换为CR1aa 50 mL/L FBS.处理组的卵裂率显著高于对照组,但囊胚率和囊胚孵化率差异不显著.实验三:对照组连续7 d均为CR1aa 50 mL/L FBS 0.1mmol/L GSH培养,处理组前3 d为CR1aa 0.1 mmol/L GSH培养,后4 d换为CR1aa 50 mL/L FBS 0.1 mmol/L GSH.处理组的卵裂率显著高于对照组,囊胚率极显著高于对照组,但囊胚孵化率差异不显著.结果表明,GSH与二步法培养系统结合相对于传统的一步法培养系统更适于牛体外受精卵的体外培养.

Study on a Two-step Culture System of Bovine Zygotes in vitro Maturated and in vitro Fertilized

In this study,we examined the effects of a two-step culture system,which use the different culture media,on early cleavage and competence of development of bovine zygotes in vitro fertilized.In experiment 1,the presumptive zygotes were randomly distributed into different treatment groups,A:the media was CR1aa 5% FBS;B:the media was CR1aa 3mg/mlBSA(0 d-3d),CR1aa 5?S(4 d-7d).The blastocyst rate of group B was significantly higher than that of group A(P<0.05),while the cleavage rate and hatched blastocyst rate were no significant differences(P>0.05).In experiment 2,A group: the media was CR1aa 5?S;B: CR1aa(0 d-3d),CR1aa 5?S(4 d-7d).The cleavage rate of group A was significantly higher than that of group B(P<0.05),while the blastocyst rate and hatched blastocysts rate were no significant differences(P>0.05).In experiment 3,A group: the media was CR1aa 5?S 0.1mM GSH;B:CR1aa(0 d-3d),CR1aa 5?S 0.1 mmol/L GSH(4 d-7 d).The cleavage rate of group B was significantly higher than that of group A(P<0.05) and the blastocyst rate of group B was more significantly higher than that of group A(P<0.01),but the hatched blastocysts rate were no significant differences(P>0.05).These results indicated that bovine zogotes in vitro maturated and in vitro fertilized can exceed the developmental block stage and have good developmental competence in the media supplemented GSH and in the two-step culture system.