Summary. The ability of antral follicular fluid obtained from sheep follicles to inhibit 3T3 fibroblasts maintained for 48 h in concentrations of fetal calf serum optimal for cell growth was examined. Addition of pooled follicular fluid to cultures resulted in a dose-dependent and reversible inhibition of [3H]thymidine incorporation. Serum from ovariectomized ewes, fetal calf serum, bovine inhibin, oestradiol-17β, testosterone, cortisol or progesterone were without effect over a range of doses. Treatment of pooled follicular fluid with charcoal–dextran did not reduce inhibitory activity which was only partly removed by heating at 85°C. Fluid obtained from large follicles (>5 mm) was more potent as an inhibitor than was fluid obtained from smaller (<5 mm) follicles. Gel chromatography of pooled fluid resolved two peaks of inhibitory activity associated with material of M r ∼ 180 000 and <10 000 respectively. No inhibitory activity was evident in fractions of serum from ovariectomized ewes chromatographed in an identical manner. These results indicate that ovine follicular fluid contains two components able to inhibit reversibly mitosis of 3T3 fibroblasts in vitro.
Keywords: fibroblast mitosis in vitro; inhibition; ovarian follicular fluid
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