Study on the Mechanism of Granulosa Cell Differentiation

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Establishment of Steroidogenic Granulosa Cell Lines Originating from WomenThe life span of granulosa cells obtained from human ovarian follicle is very short. We established new granulosa cell lines by triple transfection of primary cells with SV40 DNA,Ha-RAS gene and p53 gene. These stable cell lines proliferated rapidly and showed steroidogenic activity. cAMP stimulate progesterone production. Shifting the temperature of growth from 37C to 32C induced programd cell death. This newly established granulosa cell lines can serve as a useful tool for the study of the molecular and cellular mechanisms of inducible steroidogenesis in immortalized human granulosa cells.Effects of Thecal Cells on Apoptosis of Granulosa cellsUsing the co-culture system in which granulosa and thecal cells were maintained on the opposite surface of a collagen membrane, effects of thecal cells on apoptosis of granulosa cells was investigated. Hen theca cells stimulated hen granulosa cell apoptosis. However, such … More effect of theca cells was not detected when follicular cells were collected from bovine ovaries.Effects of Thecal Cells on Granulosa Cell DifferentiationThe possible role of thecal cells in the control of the hormonal producing activity of granulosa cells was investigated. Thecal cells reduced estradiol, inhibin and progesterone production by granulosa cells from early stage of follicular development, while augmented those by cells at late developmental stage. These results demonstrate that the thecal cells play an important role in the control of granulosa cell differentiation during ovarian follicular development.Effects of Androstenedione and Estradiol on the Meiosis-Arresting Activity of the Granulosa Cells Cultured Together with Thecal CellsThe present study explored how the meiosis-arresting activity of bovine granulosa cells is controlled by androstenedione and estradiol of the follicular fluid. This study showed that androstenedione controls the meiosis-arresting activity of the granulosa cells biphasically depending on its dose ; growing follicle fluid level of androstenedione augments the meiosis-arresting activity of granulosa cells, while atretic follicle level of androstenedione suppresses it. Estradiol does not such effect as that of androstenedione. Less