Steroidogenesis by Human Ovarian Cell Types in Culture: Influence of Mixing of Cell Types and Effect of Added Testosterone*

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Abstract
Various human ovarian cell types, such as granulosa,theca, stroma, granulosa plus theca, and granulosa plus theca plus stroma, were cultured either singly or in combination. The cultures were maintained for 6–8 days in Ham's F-12 medium enriched with 15% male monkey serum. The culture medium was changed every alternate day. In half of the cultures, testosterone (0.15 μg/ml) was present in the medium throughout the culture period. The spent culture media were frozen until assayed for estrogens and progesterone by RIA. Theca tissue secreted more estrogen than granulosa and stroma. The combined cultures of granulosa and theca secreted more estrogen compared to the sum of theca and granulosa cultured alone. Co-culture of stroma with granulosa and theca resulted in increased estrogen production for the first 2–4 days of culture only. Addition of testosterone resulted in an increase of estrogen secretion by granulosa or theca cultured either singly or in combination. Testosterone had no effect on estrogen secretion by stroma alone or when cocultured with granulosa and theca. Granulosa and theca cultured either singly or in combination but not stroma secreted detectable amounts of progesterone. Combined cultures of granulosa and theca secreted 2 to 4-fold more progesterone compared to individual granulosa and thecal cultures. The addition of testosterone to the medium brought about a 50% decrease in granulosa secretion of progesterone, whereas it brought about an increase in thecal progesterone secretion. However, testosterone had no demonstrable effect on progesterone secretion by combined cultures of granulosa and theca or granulosa plus theca and stroma.