EDS-40玻璃化冻存小鼠卵母细胞的研究

目的设计研究玻璃化液EDS一40对冻贮小鼠卵母细胞的效果。方法①按随机分组原则，分为实验组和对照组，实验组（a）采用自行设计的EDS一40玻璃化液进行玻璃化冷冻（其配方为：40％乙二醇＋18％葡聚糖），对照组（b）采用国际上已证实效果较好的EFS-40J~璃化溶液进行玻璃化冷冻，对照组（c）采用以PROH为基础的程序冷冻液进行程序冷冻。首先对EDS一40和EFS-40玻璃化溶液，行玻璃化测试；再随机选用小鼠卵母细胞，在室温25℃下分别加入EDS-40和EFS．40玻璃化溶液，玻璃化后装管并迅速投入液氮中。对照组（c）置于程序冷冻仪中进行程序化冷冻。各组均冻存2mo-3mo后，在25℃的水浴中复温后，用培养液反复洗涤后移入培养孔板培养，观察卵母细胞的成活率。将成活的卵母细胞行体外受精，观察其受精率。结果①两种玻璃化溶液能达到玻璃化效果；②EDS一40、EFS一40及对照组冷冻复温后的存活率依次为：79．34％、67．94％、56．34％；⑤EDS一40、EFS-40及对照组冻卵的受精率分别：79．17％、66．29％、51．25％。结论①EDS一40可以用来玻璃化冷冻小鼠卵母细胞；②EDS一40较EFS一40玻璃化溶液冻存小鼠卵母细胞效果更好；③玻璃化技术比传统冷冻法效果更好。

Study on the cryopreservation of mouse oocytes by the vitrification method

Objective to observation vitrification solution EDS-40 on mouse oocytes. Methods （1） randomly grouping principles, were divided into experimental group and control group, experimental group （a） a self-designed EDS-40 glass for vitrification solution （the formula is： 40% ethylene glycol ＋18% dextran ）, the control group （b） adoption of the international community has confirmed that better EFS-40 vitrification solution for vitrification in the control group （c） to adopt a PROH freezing solution-based process programming frozen. First of all pairs of EDS-40 and the EFS-40 vitrification solution, line glass test; then randomly selected mouse oocytes, at room temperature 25 ℃, respectively, joined EDS-40 and the EFS-40 vitrification solution, the glass alter-loading tubes and quickly put into liquid nitrogen. The control group （c） procedures for freezing instrument placed in the conduct of the proceedings of frozen. In each group were frozen 2 too-3 mo later, a water bath at 25 ℃ rewarming after repeated washing with culture medium after the culture plate culture moved to observe the survival rate of oocytes. The survival of the egg mother cells in vitro fertilization, the fertilization rate were observed. Results （2） The two kinds of vitrification solution to achieve glass transition effect; （1） EDS-40, EFS-40 and the control group, the survival rate after freeze thawing followed： 79.34%, 67.94%, 56.34%; （3） EDS-40, EFS-40 and the control group, the fertilization rate of frozen eggs： 79.17%, 66.29%, 51.25%. Conclusion （1） EDS-40 can be used to Vitrification of mouse oocytes; （2） EDS-40 compared with EFS-40 Vitrification of mouse oocytes cryopreserved solution better; （3） vitrification better than conventional freezing method.