小鼠卵母细胞冷冻技术研究

采用PBS作为冷冻基础液,分别用甘油和二甲基亚砜(DMSO)作为冷冻保护液,在程序化冷冻保存和玻璃化冷冻保存条件下,研究小鼠生发泡期(GV期)卵母细胞的抗冻能力。结果表明,2种冷冻方法对小鼠GV期卵母细胞解冻后形态正常率和存活率无显著影响(P>0.05)。冷冻保护剂种类对小鼠GV期卵母细胞解冻后形态正常率无显著影响(P>0.05);但对存活率有显著影响,玻璃化冷冻采用二甲基亚砜作为冷冻保护液效果极显著优于甘油(P<0.01)。以冷冻效果较好的二甲基亚砜作为冷冻保护液,采用玻璃化冷冻不同发育阶段(GV期和MⅡ期)的小鼠卵母细胞,解冻后形态正常率无显著差异(P>0.05),但存活率GV期要显著优于MⅡ期卵母细胞(P<0.05)。

Studies on cryopreservation of mouse oocytes

The anti-freezing competences of mouse GV oocytes cryopreserved by procedure and vitrification were studied after thawing. PBS was used as holding mediums and Glycerol and DMSO were used as cryopretectants. The results showed that the method of cryopreservation did not have significant effects on the rate of natural and survival mouse oocytes (P>0.05). The cryoprotectants used in vitrification freezing did not have significant effect on the rate of natural mouse oocytes (P>0.05) but on the rate of survival mouse oocytes, and DMSO was very significantly better than Glycerol in vitrification freezing (P<0.01). The anti-freezing competences of mouse GV and MⅡoocytes cryopreserved by vitrification were evaluated, which showed better results when using cryopretectant DMSO. There was no significant difference on the rate of nature between GV and MⅡoocytes (P>0.05), but the rate of survival mouse GV oocytes was significantly better than that of MⅡoocytes (P<0.05).