| To provide an alternative method for cryopreservation, and provide an new thought and way to t save the rare and endangered animals. The studies were performed on the cryopreservation of mouse morula by vitrification, the experiment was carried out as the following: research into a rapidly determination of mouse morula viability;study on influence of different freezing methods and different cryopreservation time to the development of mouse morulas ; determine traumata of cells in freezing embryo. On the base of vitrification in mice the preliminary researchs of the vitrified cryopreservation in Guizhou xiang Pig were carried on. The result of the experiment as follow:The result shows that FDA and PI can label mouse morulae which are in different viability. FDA is innocuous to embryos and PI is innocuous to embryos in intact zona pelluida.This experiment showed that whether the freezing effect stand or fall lies on vitrification cryoprotectants. When the effect of vitrification cryoprotectants is good, the effect of the container is not evident. When the effect of vitrification cryoprotectants is not good. But Freezing mouse morulae affects the integrality of their membranes, enzymatic activity and the mitochondrial distributing. But well effective freezing method little affects the integrality of their membranes, enzymatic activity and the mitochondrial distributing. Select ethylene glycol 25% EG and 25% glycerol as Vitrification Cryoprotectant , Apply plastic straw method to freezing mouse morulae, which are kept below -196°C for 1 day or 10 days or 20 days or 30 days is not affected. On the foundation of thd result, the most facile and most effective freezing method is plastic straw method using 25%Gly + 25%EG+50%mCZB.The 4-cell period embryos of Guizhou White Xiang pigs are frozen by CB+centrifugal method and frozen directly. The result showed that survival rate in vivo of vitrified morulae are high, which was similar to control. |
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