Relationship between the production of prostaglandins and progesterone by luteinizing human granulosa cells

Abstract

Luteinizing granulosa cells synthesize high concentrations of progesterone, prostaglandin (PG) E2 and PGF2α. The objective of this study was to explore the relationship between prostaglandin and progesterone output from human granulosa cells as they undergo functional luteinization in culture. Granulosa cells were partially purified from ovarian follicular aspirates and cultured at a density of 105 cells/ml in serum-supplemented DMEM:Ham's F12 medium for 0, 1 or 2 days. Cells were then switched to serum-free medium for 24 h before measuring hormone concentrations in this spent medium by specific radio-immunoassays. Over the first 3 days in culture, PGF2α and PGE2 production declined progressively by up to 82 ± 3% coincident with a 55 ± 11% increase in progesterone output. In subsequent experiments, cells were treated for 24 h on the second day of culture with either 0.01 to 10 μM meclofenamic acid or with 10 μM and 100 μM aminoglutethimide. Meclofenamic acid inhibited synthesis of PGF2α and PGE2 by up to 70 ± 9% and 64 ± 7% respectively without affecting progesterone output. Likewise, 100 μM aminoglutethimide inhibited progesterone production by 62 ± 6% without affecting concentrations of either PGF2α or PGE2. We have concluded that the progressive decline in prostaglandin production and the rise in progesterone output from luteinizing human granulosa cells occur independently of each other.