ESTRADIOL AND PROGESTERONE PRODUCTION BY CULTURED GRANULOSA-CELLS CRYOPRESERVED FROM IN-VITRO FERTILIZATION PATIENTS

Gonadotropin-stimulated steroidogenesis was studied in cultured human granulosa-lutein cells obtained from patients undergoing procedures fo r in vitro fertilization. The impact of cryopreservation on cell funct ion in vitro was studied. Granulosa cells obtained from in vitro ferti lization patients were cultured in serum-supplemented medium or cryopr eserved at -135 degrees C for 2-22 months. Fresh (unfrozen) cells (10( 5)) produced estradiol at a rate of 1320 pmol/l (over 72 h) and proges terone at about 2500 nmol/l. Estradiol production by either fresh or c ryopreserved granulosa cells in culture was unaffected by physiologica l concentrations of follicle-stimulating hormone (7 IU/l). Adding test osterone (10(-7) mol/l) to the medium increased estradiol secretion ap proximately sixfold. In contrast, progesterone production was not affe cted by follicle-stimulating hormone or testosterone. No significant d ifferences were observed in cultures of cryopreserved granulosa cells compared to cultures of unfrozen cells with respect to estradiol secre tion, the effects of follicle-stimulating hormone or testosterone on e stradiol secretion, or progesterone production. Progesterone productio n by fresh and cryopreserved cells was stimulated by human chorionic g onadotropin. These data indicate that cryopreservation offers the pote ntial to facilitate prospective studies utilizing large numbers of hum an granulosa-lutein cells in culture.