【摘要】 为获得更多具有后续发育能力的小鼠M期卵裂球,以测定细胞间期和M期的重编程因子表达情况,用不同浓度的诺考达唑(Nocodazole)对小鼠1-cell胚胎及2-cell胚胎进行了不同时间的处理,筛选出了控制卵裂球停留在M期的最佳处理浓度及处理时间。结果表明:0.10,0.15和0.20μg·m L-1的诺考达唑可有效且可逆地使小鼠1-cell胚胎或2-cell胚胎阻抑在M期;在诺考达唑去除后的1~2h,胚胎细胞重新恢复有丝分裂,完成卵裂;0.10μg·m L-1的诺考达唑处理12h可很好地将1-cell胚胎阻抑于M期,且对胚胎的后续发育影响不大;0.20μg·m L-1的诺考达唑处理18h对2-cell胚胎阻抑效果最好,而解除诺考达唑处理后3h内的胚胎发育能力略低于0.15μg·m L-1处理组,但差异不显著(p>0.05)。由于两组(0.15μg·m L-1组和0.20μg·m L-1组)对2-cell胚胎阻抑效果及发育能力两方面均差异不显著(p>0.05),因此选用0.15μg·m L-1的诺考达唑处理2-cell胚胎。用0.10μg·m L-1和0.15μg·m L-1的诺考达唑分别对小鼠的1-cell胚胎及2-cell胚胎处理12h和18h,可获得更多有发育能力的小鼠M期卵裂球。更多还原

【Abstract】 In order to obtain many more fertile mouse blastomeres at metaphase so that to mensurate the expression of reprogramming factors at interphase and metaphase, 1-cell and 2-cell mouse embryos were treated with different concentrations of Nocodazole and different time periods. Finally, optimal concentration of Nocodazole and culture time were selected. The result showed that 1-cell or 2-cell embryos could be restrained at metaphase by 0.10, 0.15 and 0.20μg·m L-1Nocodazole effectively and reversibly. The cell could recover mitosis in 1-2h after Nocodazole eliminated. 1-cell embryos could be restrained at metaphase by 0.10μg·m L-1Nocodazole with 12 h which did not affect their subsequent development. 0.20μg·m L-1Nocodazole with 18 h was better to restrain 2-cell embryos at metaphase but not more effective for subsequent development than that of 0.15μg·m L-1when Nocodazole was eliminated in 3h(p>0.05). As 0.15μg·m L-1and 0.20μg·m L-1Nocodazole did not differ significantly in restricting2-cell embryos at metaphase and for their subsequent development(p>0.05), so 0.15μg·m L-1Nocodazole was selected. 0.10μg·m L-1and 0.15μg·m L-1Nocodazole were picked up to treat 1-cell and 2-cell mouse embryos with 12 h and 18 h separately, so that many more fertile mouse blastomeres at metaphase was obtained.更多还原