In vitro effect of danazol on steroidogenesis in developing human follicles. An interim report

In order to show whether or not danazol exerts direct effects on the cells of the human ovarian follicle, follicular granulosa and thecal cells of follicles in different stages of development are currently being investigated in short-term incubation experiments and during more prolonged cell culture. Cells are isolated by free-hand dissection from follicles of 8 to 25 mm diameter, corresponding approximately to cycle day 7 to 14. Granulosa and thecal cells have so far been incubated for 2 to 4 hours in the presence or absence of hCG and/or danazol. A small number of cell cultures of granulosa cells have been performed for up to 6 days in the presence or absence of danazol, hFSH, and hLH, in various combinations. The progesterone (P), estradiol-17 beta (E) and androstenedione (A) content of the incubation or culture medium and the tissue content of cyclic AMP (cAMP) were measured. Danazol (10 micrograms/ml) inhibited thecal cAMP production in most short-term experiments but only the hCG-stimulated cAMP production of granulosa cells. Short-term production of P and E by granulosa and of A and E by thecal cells was also inhibited by danazol in certain circumstances. In tissue cultures of granulosa cells lasting up to 4 days, danazol (1 and 10 micrograms/ml) inhibited both the FSH and LH-stimulated P production. Provided that any unspecific effects of the concentrations of danazol used can be ruled out, the results seem to indicate that this substance may act directly upon steroidogenesis at the ovarian follicular level.