核抑制剂(Roscovitine)对促排周期人未成熟卵母细胞体外成熟安全性的初步研究

目的 观察核抑制剂(Roscovitine)对人未成熟卵母细胞体外成熟(in vitro maturation,IVM)安全性的影响.方法 获取促排周期的GV期未成熟卵随机分为两组,实验组(经二步法):未成熟卵母细胞先用含有200 vmol/LRoseovitine的培养液培养6 h,然后转入IVM成熟培养液培养24～36 h;对照组:直接用IVM成熟培养液培养24~36h.观察两组卵母细胞成熟情况并应用激光扫描共聚焦显微镜观察培养成熟后MⅡ期卵母细胞纺锤体和染色体的形态.结果 实验组共获未成熟卵母细胞61个,经体外成熟培养后50个卵成熟,其中39个卵可观察到纺锤体与染色体,纺锤体的形态异常者为15个,染色体异常者11个;对照组共获未成熟卵母细胞60个,经体外成熟培养后43个卵成熟,其中32个卵可观察到染色体与纺锤体,纺锤体形态异常者14个,染色体异常者11个.结论 Roscovitine能可逆性抑制体外培养的人卵母细胞的核成熟,但未对纺锤体及染色体形态产生不良影响.

The Effect of Nuclear Inhibiter(Roscovitine)on Maturation and Security in vitro in Human Immature Oocytes

Objective To investigate the effects of 200μmol/L nuclear inhibiter(Roscovitine) on maturation and security in vitro in human immature oocytes.Methods Immature oocytes were cultured with a two-step culture system.Firstly immature oocytes were cultured in maturation medium with Roscovitine for 6 h,then in maturation medium without Roscovitine for 24-36 h.Laser confocal scanning microscopy was used to examine the meiotic spindle and chromosome.The oocytes in control group were cultured in TCM-199 maturation medium.Results In experimental group 61 oocytes were observed.After in vitro maturation,50 oocytes became maturation,and 39 of them had spindle and chromosome,including 15 abnormal spindles and 11 abnormal chromosomes.In control group 60 oocytes were observed.After in vitro maturation 43 became maturation,and 32 of them had spindles and chromosomes,including 14 abnormal spindles and 11 abnormal chromosomes.Conclusion Roscovitine can reversibly inhibit nuclear maturation of human oocytes,but has no adverse effects on spindle and chromosome configuration.