氯化锶不同浓度及作用时间对小鼠孤雌胚类原核数量的影响

Effect of Different Strontium Chloride Concentrations and Action Time on the Number of Analogous Pronuclear in Mouse Parthenogenetic Embryos

目的:探讨不同浓度及作用时间的氯化锶(SrCl2)对小鼠卵母细胞孤雌激活胚胎中出现类原核数量及发育潜能的影响,建立能模拟精子效应的小鼠孤雌的激活体系。方法:分别用5 mmol/L、10 mmol/L、15 mmol/L、20 mmol/L浓度的SrCl2激活小鼠卵母细胞,观察激活胚的类原核数量及发育潜能,然后在最佳刺激浓度的SrCl2的激活液中分别处理4 h、6 h、8 h,观察类原核数目及形态,分析SrCl2浓度对小鼠卵母细胞激活效果的影响。结果:hCG注射后18 h取小鼠卵母细胞,在10 mmol/L SrCl2浓度下激活6 h,出现双原核(2PN)百分比显著高于其它浓度SrCl2处理组(P<0.01),且与小鼠体外受精6 h后的2PN率及囊胚形成率无统计学差异(P>0.05)。结论:10 mmol/L SrCl2条件下激活小鼠卵子6 h产生的孤雌胚胎中出现2PN率及发育潜能最高,提示10 mmol/L SrCl2激活处理6 h可以模拟精子对卵子的激活条件。

Objective： To explore different concentrations and action time of strontium chloride （S rC 12） on the impact of developmental potential and quantity of analogous pronuclear formation in mouse parthenoge- netic embryos, and to establish a parthenogenetic activation system that could simulate the effect of mouse sperm activation. Methods： Different concentrations including 5 mmol/L, 10 mmol/L, 15 mmol/L, 20 mmol/L of SrC12 were used to activate mouse oocytes, to select the most proper concentration of SrC12 which could induce the largest rate of activated oocytes. Mouse oocytes were treated under the most porper concentration of SrC12 for 4 h, 6 h, 8 h, respectively. Parthenogenetic activation was observed through the number of analogous pronuclear formation and developmental potential. Results： Mouse oocytes were collected 18 h after hCG injection and activated for 6 h under 10 mmol/L SrC12, the percentage of double-analogous pronuclear formation was significantly higher than other groups （P〈0.05）, the blastocyst number was also significantly higher than others （P〈0.05）, but has no significant differences compared with in vitro fertilization （IVF） 6 h embryo group （P〉0.05）. Conclution： When mouse oocytes activating for 6 h under 10 mmol/L SrC12, double-analogous pronuclear proportions and developmental potential of the parthenogenetic embryos were the highest, suggesting that 10 mmol/L SrC12 combined 6 h activation process can simulate sperm activation conditions.