In Vitro Differentiation of Bovine Granulosa Cells Obtained from Small Antral Follicles

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The present study was undertaken to investigate whether bovine granulosa cells obtained from small antral follicles could differentiate into luteal-like cells in vitro. Granulosa cells obtained from small antral follicles (3-5 mm in diameter) were cultured up to 9 days in the presence or absence of insulin (2 μg/ml), forskolin (10 μM) and insulin (2 μg/ml) plus forskolin (10 μM). On days 1, 3, 5, 7 and 9 of culture, spent media were harvested for hormone assays. Progesterone production in the granulosa cells incubated with insulin plus forskolin increased continuously during the culture period. On the other hand, estradiol-17β production in the granulosa cells sharply decreased on day 3, and then continued to decrease. The size of granulosa cells cultured with insulin plus forskolin for 9 days was significantly larger than that of granulosa cells at the beginning of culture (P<0.001), and the size was similar to that of large luteal cells. Collectively, these data suggest that bovine granulosa cells obtained from small antral follicles can differentiate into large luteal-like cells during culture in the presence of insulin and forskolin.
日本繁殖生物学会の論文