Steroid and plasminogen activator production by cultured rat granulosa cells in response to hormone treatment.

Granulosa cells isolated from immature, DES-primed female rats were incubated in medium-199 plus 10% chicken serum with addition of FSH, or testosterone, or both. Cultures were incubated at 37 degree C for 7 days; medium samples were taken daily and analyzed for steroids and plasminogen-activator production. Only cultures containing FSH + testosterone produced significant amounts of both estradiol and progesterone after 2 days of incubation. The rate of estradiol production increased steadily up to the 4th day and then leveled off; the production of progesterone reached a maximum around the 3rd day, and then declined rapidly afterward. FSH alone was able to stimulate plasminogen activator production at the first day. Cultures with FSH + testosterone produced an additional peak of plasminogen activator activity at the 4th day. Plasminogen-activator production is thus not correlated with steroidogenesis in a simple way. We conclude that the granulosa cell require the presence of both FSH and testosterone at the beginning of incubation for normal response. Delayed addition of either hormone, or both, to the culture causes damage to the cells ability to produce normal responses to hormone treatment.