A granulosa cell-inhibitory factor (GCIF) of low molecular mass from bovine follicular fluid inhibits the proliferation of bovine granulosa cells in vitro and the growth of large follicles in rats in vivo. The present study examined the ability of GCIF to modulate the effects of FSH, epidermal growth factor (EGF), insulin-like growth factor I (IGF-I) and androstenedione on the proliferation of bovine granulosa cells and on aromatase activity in vitro. Granulosa cell proliferation was assayed by

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... counting haemocytometric cells and by measuring the incorporation of [3H]thymidine into acid-precipitable material. Assay of aromatase activity was based on the conversion of [3H]androstenedione to [3H]H2O. FSH, androstenedione, EGF and IGF-I all stimulated (P < 0.01) granulosa cell proliferation; however, the addition of GCIF reduced (P < 0.01) cell proliferation in their presence. In the case of EGF, the addition of GCIF almost abolished the stimulatory response. FSH and IGF-I, but not EGF, stimulated (P < 0.01) aromatase activity of granulosa cells. The stimulatory effect of IGF-I was decreased by GCIF. The inhibitory effects of GCIF indicate that it may play a significant role in regulating the effects of intraovarian growth factors on granulosa cells and the growth of follicles.