GRANULOSA CELL FUNCTION DURING FOLLICULOGENESIS

卵泡的生长和发育依赖于 卵泡内有大量的颗粒细胞。系统的研究 颗粒细胞的生长和分化一直很困难 由于同一卵泡内颗粒细胞的差异。 可以分离出功能不同的颗粒细胞群 用Percoll梯度法从未成熟大鼠的卵巢中提取 己烯雌酚(DES)诱导卵泡生长和妊娠母马的实验研究 血清促性腺激素(PMSG)诱导卵泡生长和分化。 两种颗粒细胞群被证明是通过流动而存在的 细胞学和EM研究。DES和PMSG处理的大鼠颗粒细胞 将使用EPICS 5流式细胞仪和Percoll进行分离 渐变。电池将以光和电子为特征 显微镜下，FSH和LH受体的存在以及FSH诱导的雌二醇和 黄体酮的产生。卵巢的一种浓缩组合 卵泡的大小和颗粒细胞的分离有望产生 不同生长和发育阶段的颗粒细胞群 差异化。使用DES和PMSG免疫的未成熟大鼠正在进行 以具有成本效益的方式制定实验程序 最低限度的动物使用和所获得的结果将在成人身上得到验证 骑自行车的老鼠。为了获得足够数量的颗粒细胞 不同生长发育阶段和分化利用的进一步研究 在最小数量的动物中，通过将 将尝试SV40病毒的大T抗原来产生稳定的细胞 台词。这样的转基因研究在我们手中已经成功地使用了 牛肾上腺细胞，细胞在其中保留其预转染 特点。纯化的颗粒细胞群具有不同的 生长和分化特征，届时将进行详细研究 关于生长和分化的调节，如 生长因子、雌二醇、cAMP类似物和卵泡刺激素单独或组合 在无血清限定的培养液中。雄激素在分期中的作用 对芳香酶的依赖刺激或抑制也将进行研究。 这些研究有望为以下因素提供新的见解： 在此过程中导致健康的排卵卵泡或闭锁卵泡 卵泡发生的过程。

The growth and development of ovarian follicles is dependent upon the population of granulosa cells present in the follicle. A systematic study of the growth and differentiation of granulosa cells has been difficult because of the differences in granulosa cells within the same follicle. Functionally different populations of granulosa cells can be separated using Percoll gradients from the ovaries of immature rats treated with diethyl stilbestrol (DES) to induce follicular growth and pregnant mare's serum gonadotropins (PMSG) to induce follicular growth and differentiation. Two populations of granulosa cells gave been shown to exist by flow cytometry and EM studies. Granulosa cells from DES and PMSG treated rats will be separated by the use of an EPICS 5 flow cytometer and Percoll gradients. The cells will be characterized by light and electron microscopy, presence of FSH and LH receptors and FSH induced estradiol and progesterone production. A combination of the enrichment of ovarian follicles by size and granulosa cell separation is expected to yield populations of granulosa cells in various stages of growth and differentiation. The use of the DES and PMSG primed immature rats is being made to work out experimental procedures in a cost effective manner with minimal animal use and the results obtained will be verified in the adult cycling rat. In order to obtain sufficient numbers of granulosa cells for further studies in different stages of growth and differentiation with use of a minimal number of animals, immortalization by transfection with the large T antigen of the SV40 virus will be attempted to generate stable cell lines. Such transfection studies have been successful in our hands using bovine adrenal cells in which the cells retained their pretransfection characteristics. With purified granulosa cell population with different growth and differentiation characteristics, detailed studies will then be done on the regulation of growth and differentiation by agents such as growth factors, estradiol, cAMP analogues, and FSH singly or in combination in a serum free defined medium. The effect of androgens on the stage dependent stimulation or inhibition of aromatase will also be studied. These studies are expected to provide new insights into the factors that lead to healthy ovulatory follicles or atretic follicles during the process of folliculogenesis.