Estimation of inhibin-like activity in spent medium from rat ovarian granulosa cells during long-term culture.
Granulosa cells were isolated from antral follicles of intact, adult female rats by treatment with EGTA and hypertonic sucrose, and were kept in culture for 1, 15 or 37 days. The amount of inhibin-like activity in spent rat granulosa-cell media (rGCCM) was assessed by using a bioassay for inhibin (measurement of the unstimulated release of FSH and LH by pituitary cells in culture). After 1 day in culture, inhibin-like activity was detected in rGCCM of 0.016-0.8 X 10(5) cultured granulosa cells per dish. In the bioassay, maximal suppression of FHS occurred when 0.8 X 10(5) granulosa cells were plated. With this or a larger number of cells in long-term cultures, pooled rGCCM, collected up to 37 days after plating, suppressed levels of FSH (and not of LH) in the inhibin bioassay in parallel with the suppression found after addition of charcoal-treated bovine follicular fluid (bFF), which served as a standard. The amount of inhibin-like activity of rGCCM (relative to the amount present in bFF, which was given a potency of 1) ranged between 0.05 and 0.90 X 10(-3). Release of inhibin-like activity could not be estimated on day 4, and was low from days 29 to 37 in culture. Except after 1 day of culture, no substantial amounts of progesterone were detected in the media. It is concluded that granulosa cells collected from adult, intact female rats have and retain the capacity to secrete inhibin-like activity in vitro, under basal conditions in long-term cultures.
D000818:Animals; D001681:Biological Assay; D002478:Cells, Cultured; D004533:Egtazic Acid / Q000494:pharmacology; D005260:Female; D005640:Follicle Stimulating Hormone / Q000378:metabolism; D006107:Granulosa Cells / Q000032:analysis*; D007265:Inhibins / Q000032:analysis*; D007986:Luteinizing Hormone / Q000378:metabolism; D010902:Pituitary Gland / Q000187:drug effects; D051381:Rats; D011919:Rats, Inbred Strains; D013395:Sucrose / Q000494:pharmacology; D013997:Time Factors
D004533:Egtazic Acid; D013395:Sucrose; D007265:Inhibins; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone