人胎儿窦前卵泡体外发育的研究  

Development of Human Fetal preantral Follicles in Vitro

作　　者：吴际[1] 张丽珠[1] 刘平[1] 高荣莲[1] 

机构地区：[1]北京医科大学第三临床医学院妇产科

出　　处：《中华妇产科杂志》1998年第9期520-523,I015,共5页Chinese Journal of Obstetrics and Gynecology

摘　　要：目的了解人胎儿窦前卵泡体外发育、其卵母细胞成熟的过程和特征，影响因素及其机理。方法对来自３６例水囊中期引产、胎龄为２０～２８周的胎儿卵巢，进行体外培养，采用Ｎｏｒｔｈｅｒｎ印迹杂交及巢式聚合酶链反应技术等方法，观察卵泡细胞甲胎蛋白（ＡＦＰ）ｍＲＮＡ、ｂｃｌ－２和野生型ｐ５３ｍＲＮＡ的变化。结果（１）人胎儿窦前卵泡在体外能生长发育，随培养时间的增加，卵泡直径增大，有部分卵泡发生退化。经表皮生长因子（ＥＧＦ）或胰岛素样生长因子－１（ＩＧＦ－１）作用并经最后促成熟，卵母细胞达到成熟，成熟率ＩＧＦ－１作用为最高（Ｐ＜０．０５）。（２）ＡＦＰ基因ｍＲＮＡ在胎儿卵泡的细胞培养的５、１５、２５、３５天均有表达，而在４５天则无表达。（３）与对照比，绝经期促性腺激素（ｈＭＧ）、卵泡液、ＩＧＦ－１及ＥＧＦ作用时ｂｃｌ－２基因ｍＲＮＡ表达分别增高１７．６％、２２．４％、４７．１％和３７．５％，野生型ｐ５３基因ｍＲＮＡ表达分别降低１９．８％、２２．１％、４２．６％和３１．６％。结论（１）胎儿窦前卵泡在体外能发育其卵母细胞能成熟，ＩＧＦ－１对其促成熟作用最强。发育过程中特征为：整个发育过程中未形成明显的卵泡腔，于发育的前３５天有ＡＦ?Objective To study oocyte maturation of human fetal preantral follicles in vitro, the effects of different factors on oocyte maturation. Methods Preantral follicles were obtained from fetal ovaries from termination of 20～28 weeks gestation. Factors involved in the development, maturation of fetal preantral follicles were studied in tissue and cell culture by means of Northern blot, and RTPCR (nested). Results (1) Preantral follicles obtained from fetal ovary increased in size to more than 120 μm in diameter without antrum formation and with increasing time the preantral oocytes became mature with extrusion of 1st polar body, insulinlike growth factor 1(IGF1) had a more marked effect on oocyte maturation, as compared with epidermal growth factor (EGF). (2) Expression of alpha fetoprotein mRNA occurred in fetal follicles on day 5, 15, 25, 35 of culture, and became negative on day 45 of culture. (3) Medium consisted of ①human menopasual gonadotropin (hMG), ② follicular fluid, ③ IGF1, ④ EGF increased the expression of bcl2 mRNA by 17.6%, 22.4%, 47.1% and 37.5% respectively in fetal follicular cells, and downregulated the expression of p53 mRNA by 19.8%, 22.1%, 42.6% and 31.6% respectively. Conclusions (1) The development and maturation of fetal preantral follicles in vitro differed from those in vivo. (2) Expression of bcl2 mRNA increased and expression of wildtype p53 mRNA decreased during follicular development and maturation.

关 键 词：囊状滤泡 体外发育 卵母细胞 窦前卵泡 

分 类 号：R321－33[医药卫生—人体解剖和组织胚胎学]