Studies On Embyo Splitting And On Culture In Vitro Of The Splitted Embryos Of Mice

In the experiment, the FSH+LH combinations were adopted to superovulate KM mice to ascertain the proper phase to gain morulae and blastocysts, three ways were adopted to split the morulae and blastocysts, demi-embryos were cultured in D-PBS, mTCM199, CZB, mM16 added with FCS or MS of different concentration, and transplantation of the blastocysts derived from demi-embryos was performed. The results of the studies were as follows.1. In the experiment, the FSH+LH combinations of 4 doses were adopted to superovulate KM mice (FSH was injected by IP 4 times degressively, and LH by IP once.). It indicated that 9IU FSH+9IU LH was the best dose in which 30.71 good embryos were got from each mouse averagely and it showed the effect of 9IU FSH+9IU LH was significantly better than that of the other three groups.2. With the above scheme, the result of gaining embryos from mice with vaginal plug at different time points indicated that morula phase was mainly between 66h and 81h (especially between 69h and 78h) after LH injection and that blastocyst phase was mainly between 84h and 87h after LH injection.3. Three ways were adopted to split the morulae and blastocysts. The result indicated that achievable ratios of splitting by micro-manipulator with micro-blade or with micro-glass-needle (morula 93.0%, 90.0%, 91.5%, 91.1%; blastocyst 90.5%, 93.0%, 90.2%, 94.0%) were higher than that of splitting by hand with micro-glass-needle (morula 87.7%, 88.2%; blastocyst 87.7%, 89.6%). However, there was no significant difference between them.4. In the experiment, demi-embryos were cultured in D-PBS, mTCM199, CZB, mM16 added with FCS or MS of different concentration. The successful developmental ratio of demi-morulae was...