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Human Chorionic Gonadotropin Stimulation of Estradiol Production and Androgen Antagonism of Gonadotropin-Stimulated Responses in Cultured Human Granulosa-Luteal Cells^*

Reproductive Biology Section, Department of Obstetrics and Gynecology yale University School of Medicine New Haven,Connecticut 06510

Address request for reprints to:Dr.Mary Lake Polan, Reproductive Biology Section, Department of Obstetrics and Gynecology, Yale University School of Medicine, 333 Cedar Street, New Haven, Connecticut 06510.

Baseline and gonadotropin-stimulated estradiolproduction were examined in long term cultures of human granulosa-luteal cells isolated from women undergoing in vitro fertilization. Estradiol production declined by 70% during the first 48 h in culture and was minimally stimulated by the addition of hCG to the culture medium. During subsequent culture from 48–120 h estradiol production was significantly increased over control levels by hCG concentrations greater than 0.1 IU/ml.Incubation with testosterone stimulated estradiol production 100-fold in the presence and absence of gonadotropin hCG(0.01–10 IU/ml) stimulated a 3- to 13-fold increase in progesterone production. However, at hCG concentrations greater than 1 IU/ml, coincubation with testosterone (10^–7 M) significantly inhibited progesterone production. Dihydrotestosterone also inhibited progesterone production, but to a lesser extent than testosterone.

Freshly isolated granulosa-luteal cells specifically bound small amounts of [¹²⁵I]hCG (<l,000 cpm/105 cells). Glycine buffer wash was shown to reversibly remove more than 88% of bound hCG and, in freshly isolated cells, increased [¹²⁵I]hCG binding by 100%. In 5-day cultures, specific [¹²⁵I]hCG binding nearly doubled from 52,000 cpm/10⁵ cells in control cultures to 87,000 cpm/10⁵ cells in cultures treated with hCG (0–5 IU/ml). At the highest concentration of hCG (5 IU/ml), testosterone (10^–7 M) significantly inhibited the amount of [¹²⁵I] hCG specifically bound. In summary, estradiol production in long term cultures of granulosa-luteal cells appears to be gonadotropin dependent. In addition, the presence of testosterone (10^–7 M) antagonizes hCG-stimulated progesterone and LH receptor production by these cells.

^* This work was supported by USPHS Grants HD-05927 and HD10718.

Received September 11, 1985.

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