| 饶金鹏,邱枫,金敏.解冻液I温度设定的不同对小鼠卵母细胞玻璃化冻融效果的影响[J].浙江中西医结合杂志,2016,26(9): |
| 解冻液I温度设定的不同对小鼠卵母细胞玻璃化冻融效果的影响 |
| The effects of setting temperature of thawing medium I to 37℃ or 22℃on mouse oocytes vitrification |
| 投稿时间：2016-06-10 修订日期：2016-06-17 |
| DOI： |
| 中文关键词: 解冻液I 温度 小鼠卵母细胞 玻璃化冷冻 复苏率 |
| 英文关键词:thawing medium I temperature mouse oocytes vitrification survival rate |
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| 中文摘要: |
| 『』 目的 探讨预热至37℃的解冻液I和预热至室温(22℃)的解冻液I对小鼠卵母细胞玻璃化冻融效果的影响。 方法 对6周龄昆明系雌鼠进行促排卵,脱去颗粒细胞后选取形态良好的MII期卵母细胞,用含不同浓度DMSO+EG的磷酸盐缓冲液(5%DMSO+5%EG,10%DMSO+10%EG,20%DMSO+20%EG+30%蔗糖)对其进行玻璃化冷冻,储存2天后对小鼠卵子进行解冻复温。将卵子随机分为2组,一组投入预热至37℃的解冻液I(含30%蔗糖的磷酸盐溶液),另一组投入预热至室温(22℃)的相同浓度的解冻液I中,再分别移至预热至室温(22℃)的解冻液II,III和IV(含20%,10%,5%蔗糖的磷酸盐溶液),随后对两组卵母细胞的复苏率进行统计比较。 结果 共计对119枚小鼠卵子进行玻璃化冻融,其中61枚置于预热至37℃的解冻液I,有52枚存活良好,复苏率为82.25%(52/61),其余58枚置于预热至室温(22℃)的解冻液I进行解冻,有23枚存活良好,复苏率为39.66%(23/58),37℃组的复苏率显著高于室温(22℃)组(P<0.01)。 结论 虽然降低平衡时的温度可以减少高浓度玻璃化冷冻保护剂对卵子的毒性作用,但较快的升温速度可以帮助卵子快速通过-120℃～-35℃的危险阶段,而37℃的解冻液I环境较之室温(22℃)能更好的加速这一过程,避免重结晶对细胞的致命损伤。 |
| 英文摘要: |
| 『』Objective To evaluate the effects of thawing medium I preheating to 37℃ and 22℃ on mouse oocytes vitrification . Method To get cumulus-oocyte-comlexes(COCs), the female mice aged six weeks were induced to superovulate. After the granulosa cells of COCs were taken off in hyaluronic acid, the mature oocytes were freezed by using phosphate buffer containing different concentrations of DMSO+EG((5% DMSO+5% EG,10% DMSO+10% EG,20% DMSO+20% EG+30% sucrose). After 2 days, the frozen oocytes were randomly allocated to two thawing groups using thawing medium I (phosphate buffer containing 30% of sucrose) preheating to 37℃ and room temperature (22 ℃) respectively, and then to thawing mediumⅡ,Ⅲ and Ⅳ.The survival rates of the two groups were calculated. Results The survival rate of thawing medium I preheating to 37℃ is 82.25%(52/61) which is significantly higher than the 22℃ group 39.66%(23/58), P<0.01. Conclusions Though balance in lower temperature can reduce the toxic effects of high concentrations of vitrification cryoprotectants on the oocytes, a faster heating rate can help the oocytes quickly pass the dangerous phase of - 120 ℃ ~ -35 ℃. 37 ℃ thawing medium I environment could speed up this process better than the room temperature (22 ℃) one, and could avoid the recrystallization which cause fatal damage to the cells. |
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