细胞松弛素B和解冻方法对玻璃化冷冻小鼠卵母细胞的影响

Effects of Cytochalasin B and Thawing Program on Vitrification of Mouse Mature Oocytes

【目的】探讨细胞松弛素B和解冻程序对玻璃微细管法(GMP)玻璃化冷冻小鼠卵母细胞的影响。【方法】以小鼠MII期卵母细胞为模型,研究冷冻前细胞松弛素B预处理、不同解冻程序对小鼠卵母细胞冷冻效果的影响;随后对经GMP玻璃化冷冻的卵母细胞直接进行培养以检测冷冻是否诱发孤雌发育。【结果】与对照组相比,细胞松弛素B预处理的卵母细胞存活率、受精率、卵裂率及囊胚率没有显著差异(89.3%vs91.3%,44.0%vs40.4%,30.0%vs27.7%,4.0%vs6.4%;P>0.05);采用连续浓度梯度递减解冻法的受精率明显高于间断浓度梯度递减解冻法(57.4%vs40.4%;P<0.05),且前者的卵裂率和囊胚发育率也相对较高(40.2%vs27.7%,14.5%vs6.4%;P>0.05)。冷冻复苏后卵母细胞的孤雌发育率稍高于未经冷冻的卵母细胞(17.1%vs3.2%;P>0.05)。【结论】细胞松弛素B预处理对MII期小鼠卵母细胞的GMP玻璃化冷冻保存效果没有影响;采用连续浓度梯度递减解冻法能明显提高复苏后卵母细胞的体外发育能力。

[Objective] Effects of cytochalasin B and thawing program on glass micropipette （GMP） vitrification of mouse mature oocytes were investigated. [Methods] The effects of cytochalasin B pre-treatment and thawing program on viability and in vitro development of matured mouse oocytes were investigated in the present study. The thawing oocytes were directly cultured to explore whether vitrification caused parthenogenetical development. [Results ] Compared to the controled group, The survival rate, fertilization rate, cleavage rate, and blastocyst rate of cytochalasin B pre-treatment were not significantly different （89.3% vs 91.3%, 44.0% vs 40.4%, 30.0% vs 27.7%, 4.0% vs 6.4%; P 〉0.05）; But the fertilization rate using the thawing program of consecutive-decrease concentration were significantly higher than using the discontinuous-decrease concentration （57.4% vs 40.4%; P〈 0.05）, the cleavage rate and blastocyst rate were also higher （40.2% vs 27.7%, 14.5% vs 6.4%）, but were not statistically significant （P 〉0.05）. Vitrification were easy to induce thawing mouse oocytes producing parthenogenetical development, but not significantly different when compared to the no-vitrification group （17.1% vs 3.2%, P 〉0.05）. [Conclusions] The effects of GMP vitrification of mouse mature oocytes were not improved if pre-vitrification treatment with cytochalasin B, but the thawing program of consecutive-decrease concentration method were helpful to improve the in vitro developmental ability of mouse oocytes.