RECOMBINANT HUMAN GONADOTROPINS STIMULATE STEROID AND INHIBIN PRODUCTION IN HUMAN GRANULOSA-CELLS

Objective: Recently pure gonadotropins have become available through r ecombinant technology In parallel with ongoing clinical trials it is i mportant to examine the effects of these new gonadotropin preparations in experimental studies in human granulosa cells. In the present stud y the effects of recombinant FSH (rFSH) and LH (rLH) on steroid and in hibin production were examined in human granulosa cells in culture. Pa tients and methods: Granulosa cells were obtained during the follicula r phase of the menstrual cycle in seven women undergoing gynecological laparotomy and from follicles in stimulated cycles in women undergoin g oocyte retrieval in connection with in vitro fertilization/embryo tr ansfer. The granulosa cells were cultured in modified Medium 199 conta ining 1% fetal bovine serum for 4-8 days with and without hormones. Me dia were changed on alternate days and stored at -20 degrees C until a nalyzed for estradiol, progesterone and inhibin. Results: Granulosa ce lls from natural cycles were highly responsive to rFSH which caused a dose-related (rFSH 0.1 to 100 ng/ml) increase in estradiol and progest erone accumulation. The maximal stimulatory effect was reached with a concentration of rFSH between 1 and 10 ng/ml. Granulosa cells from sti mulated cycles responded highly to rLH in terms of increased progester one production during the whole culture period. A maximal stimulatory effect was observed with rLH at a concentration of 0.1 ng/ml. Both typ es of granulosa cells responded to recombinant gonadotropins in terms of increased inhibin production. Conclusions: The present study demons trates that granulosa cells from human ovarian follicles are highly re sponsive to recombinant gonadotropins as demonstrated by increased ste roid and inhibin production.