Hela细胞MPF对未成熟卵母细胞的体外促成熟作用分析  

Assay of promoting-maturation activity of M-phase promoting factor(MPF) from Hela cell

作　　者：彭佑共[1] 杨罗艳[1] 

机构地区：[1]中南大学湘雅二医院泌尿外科,湖南长沙410011

出　　处：《医学临床研究》2008年第9期1624-1627,共4页Journal of Clinical Research

摘　　要：【目的】研究Hela细胞M期促进因子MPF对未成熟卵母细胞的体外促成熟作用的生物活性,并观察单个成熟卵母细胞的中期染色体。【方法】应用秋水仙胺将Hela细胞阻滞在有丝分裂的中期(meta-phase),从中提取MPF蛋白;体外培养基分为无激素组、有激素素组、注射组,将MPF提取物通过显微注射,分别导入3组培养基中的昆明小白鼠未成熟卵母细胞中,观察未成熟卵母细胞发生GVBD(生发泡破裂)、或者排出极体的数量,并对3组两两之间发生GVBD或者排出极体的情况,作统计分析;以及制备并观察成熟卵母细胞的中期相染色体。【结果】经过统计分析说明,发生GVBDD的情况,在注射组与无激素组之间差异有显著性(P<0.05);排出极体的情况,在注射组与无激素组之间,以及注射组与激素组之间差异有显著(P<0.005,P<0.05)。已经观察到了部分成熟卵母细胞的中期染色体。【结论】MPF提取物具有诱导小白鼠未成熟卵母细胞在体外提前恢复减数分裂的生物活性。[Objective]To assay the promoting-maturation activity of MPF to induce immature oocytes to mature in vitro and to observed the metaphase chromosomes of the single mature oocytes. [Methods]Hela cells were blocked at mitotic metaphase by colcemid in order to extract the proteins containing MPF. The culture medium was individed into 3 groups, non-hormone group（without hormone）, hormone group（with hormone）, injecting-group（without hormone）. MPF extracts were microinjectied into immature oocytes of Kunming mouse in the injecting-group medium. Observation and statistic analysis of the cell number with GVBDs（germinal vesicle breakdown） or extrued polar bodies were made among the three group. The metaphase chrosomes of mature oocytes were prepared and observed. [Results]The significant difference of the percent of GVBDs was demonstrated between non-hormone group and injectied-group by statistics （ P 〈0.05）, The percentage of extrued polar bodies in injected group was significantly higher than non-hormone group and hormone group （53.4% vs 31.08%, 33.3 %）, The significant difference was demonstrated between the injectied-group and non-hormone group ,or hormone group by statistics （ P 〈0. 001, P 〈0. 005）. And the metaphase chromosomes of the single mature oocytes were observed. [Conclusion]The results indicated MPF extract possess the property that can induce immature oocytes to reinitiate meiosis.

关 键 词：卵母细胞 HELA细胞 成熟促进因子 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]