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Journal of Clinical Endocrinology & Metabolism, Vol 75, 911-917, Copyright © 1992 by Endocrine Society
ARTICLES |
JH Kim, MM Seibel, DT MacLaughlin, PK Donahoe, BJ Ransil, PA Hametz and CJ Richards
Faulker Center for Reproductive Medicine, Faulkner Hospital, Deaconess/Harvard Surgical Program, Boston, Massachusetts.
This study was undertaken to determine if Mullerian-inhibiting substance (MIS) could block basal and/or epidermal growth factor (EGF)- induced proliferation and progesterone production by cultured human granulosa-luteal cells. Cells from follicles of individual patients were pooled, counted, and aliquoted into Ham's F-10 medium containing 10% MIS-free female fetal calf serum at 37 C in 95% air and 5% CO2. After assessing viability, cells were counted on days 4, 8, 12, and 16 of culture. EGF was added every other day at 0.2, 2, and 20 ng/mL beginning on culture day 4. The greatest stimulatory effect of EGF on cell proliferation was observed at 20 ng/mL on days 12 and 16. EGF increased progesterone production per cell after 4 days exposure, but this effect was lost after 8 days. Granulosa-luteal cells were cultured with 0.2, 2, and 20 ng/mL immunoaffinity purified recombinant human MIS (rhMIS) or conditioned medium from Chinese hamster ovary cells transfected with the human MIS gene, beginning on culture day 4. rhMIS demonstrated its greatest inhibitory effect on cell proliferation at 20 ng/mL on day 16. The rhMIS decreased progesterone production per cell after 4 days exposure, but only in the higher doses. Maintaining EGF at 20 ng/mL and varying rhMIS yielded significant reduction in EGF- mediated proliferation and progesterone production per cell at 2 and 20 ng/mL rhMIS. These experiments demonstrate rhMIS inhibits basal and EGF- stimulated human granulosa-luteal cell proliferation and progesterone production.
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