Study for Effects of Ovarian Thecal Cells on Meiosis-Arresting Activity and Differentiation of Granulosa Cells

我们已经成功地开发了一种共培养系统，其中卵巢颗粒细胞和卵泡膜细胞生长在胶原膜的相对侧。使用这种共培养系统，我们以前已经证明，这两种卵泡细胞类型调节其增殖，形态，结构和functions.In第一系列的研究，我们已经检查了使用这种培养系统的颗粒卵泡膜细胞通信卵母细胞成熟的效果。牛卵丘-卵母细胞复合物与颗粒细胞孵育，有或没有卵泡膜细胞。评估了生殖囊泡破裂的时间过程。当与颗粒细胞和卵泡膜细胞一起培养时，与单独与颗粒细胞一起培养时相比，生殖囊泡破裂显著延迟。当卵丘-卵母细胞复合体与FSH或低剂量雄烯二酮处理的颗粒细胞和卵泡膜细胞孵育时，分解进一步延迟。而对胚芽囊泡破裂的影响则未被检测到。 ...更多信息 第二部分研究了卵泡膜细胞对体外培养的早、中、晚期卵泡颗粒细胞激素分泌活性及超微结构的影响。卵泡膜细胞在卵泡发育早期减少颗粒细胞的激素分泌，而在卵泡发育晚期增加颗粒细胞的激素分泌。与卵泡膜细胞共培养的颗粒细胞的形态学分析显示，与单独培养的细胞相比，来自小卵泡的细胞的溶酶体聚集、内质网和线粒体收缩减少，但来自成熟卵泡的细胞的这些细胞分化特征增强。这些结果表明，卵泡膜细胞分泌的因子抑制未成熟颗粒细胞的分化，而促进成熟的颗粒细胞的分化，并建议卵泡膜细胞在控制卵巢卵泡发育过程中的颗粒细胞分化中发挥重要作用。少

We have successfully developed a co-culture system in which ovarian granulosa and thecal cells are grown on opposite sides of a collagen membrane. Using this co-culture system, we have previously demonstrated that the two follicular cell types reciprocally modulate their proliferation, morphology, structure and function.In the first series of the study, we have examined the effect of granulosa-theca cell communication on oocyte maturation using this culture system. Bovine cumulus-oocyte complexes were incubated with granulosaa cells, with or without theca cells. The time course of germinal vesicle breakdown was assessed. When cultured with granulosa and theca cells, germinal vesicle breakdown was significantly delayd compared when cultured with granulosa cells alone. Breakdown was further delayd when cumulus-oocyte complexes were incubated with FSH- or low-dose of androstenedione-treated granulosa cells with theca cells. While, such effects on germinal vesicle breakdown was not detecte … More d when granulosa cells were incubated with high dose of androstenedione or estradiol.In the second series of the study, the influence of thecal cells on the hormonal producing activity as well as the ultrastructure of granulosa cells from early-, mid-and late-developmental stage follicles in vitro was investigeted. Thecal cells reduced hormonal production by granulosa cells from early stage of follicular development, respectively, while augmented that by cells at late developmental stage. Morphological analysis of granulosa cells co-cultured with thecal cells shows a decrease in the clustering of lysosomes, shrinkage of endoplasmic reticulum and mitochondria of cells from small follicles but an enhancement of these cytodifferentiative features in cells from mature follicles when compared to the cells cultured alone. These results demonstrate that the thecal cells secrete factor (s) which inhibits the differentiation of the immature granulosa cells while promoting that of the matured ones and suggest that the thecal cells play an important role in the control of granulosa cell differentiation during ovarian follicular development. Less