High survival of mouse oocytes using an optimized vitrification protocol.
High survival of mouse oocytes using an optimized vitrification protocol.
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使用优化的玻璃化冷冻方案提高小鼠卵母细胞的存活率
DOI：
10.1038/srep19465
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发表时间：
2016-01-19
影响因子：
4.6
通讯作者：
Liang CG
中科院分区：
文献类型：
--
作者：
Zhou CJ;Wang DH;Niu XX;Kong XW;Li YJ;Ren J;Zhou HX;Lu A;Zhao YF;Liang CG
玻璃化冷冻法已广泛应用于超低温保存。然而，这种方法对哺乳动物卵母细胞的有效性可以通过优化该过程的每个步骤来提高。在本研究中，我们测试了不同的几个关键参数的影响，以确定最有效的方案，小鼠卵母细胞玻璃化冷冻。我们发现含有乙二醇和二甲亚砜的冷冻保护剂加20%胎牛血清产生的卵母细胞存活率、受精率和囊胚形成率最高。卵母细胞在玻璃化冷冻液和解冻液中冷冻的时间和温度对卵母细胞的存活率有影响。卵母细胞周围卵丘细胞的存在和解冻卵母细胞在Toyoda-Yokoyama-Hosoki培养基中的孵育也增加了卵母细胞的存活率。开拉吸管法和尼龙圈法比微滴法更有效。最后，与未改进的方法相比，这些改进的方法的组合导致更好的纺锤体形态。这些结果表明，小鼠卵母细胞玻璃化冷冻的结果可以通过适当的冷冻保存方法的组合，这可以应用于未来的临床研究与人类卵母细胞。
The method of vitrification has been widely used for cryopreservation. However, the effectiveness of this method for mammalian oocytes could be improved by optimizing each step of the process. In the present study, we tested the effects of varying several key parameters to determine the most effective protocol for mouse oocyte vitrification. We found that cryoprotectant containing ethylene glycol and dimethylsulfoxide plus 20% fetal calf serum produced the highest rates of oocyte survival, fertilization and blastocyst formation. The duration and temperature of oocyte exposure to vitrification and thawing solutions influenced survival rate. The presence of cumulus cells surrounding oocytes and the incubation of thawed oocytes in Toyoda-Yokoyama-Hosoki medium also increased oocyte survival. Open pulled straw and nylon loop methods were more effective than the mini-drop method. Finally, the combination of these improved methods resulted in better spindle morphology when compared to the unimproved methods. These results demonstrate that the outcomes of mouse oocyte vitrification can be improved by a suitable combination of cryopreservation methods, which could be applied to future clinical research with human oocytes.