Influence of gonadotropins on steroidogenesis by large follicle buffalo granulosa cells under in vitro serum-free conditions

Abstract

The study was made to examine the effects of FSH and LH on estradiol–17β and progesterone production by large follicle derived buffalo granulosa cells cultured under completely serum-free conditions. Granulosa cells (3×105) from large (>9 mm diameter) follicles were cultured for 24 h in 48-well plates coated with 3.3μg/cm2 fibronectin in DMEM: Nutrient mixture F-12 Ham (1:1 ratio) supplemented with 10-7 M androstenedione, 5 mg/ml human apo-transferrin and 0.1% BSA, in the presence or absence of FSH or LH (1 to 64 ng/ml each). FSH had no effect on estradiol-17β output but at 4 and 8 ng/ml increased progesterone secretion by granulosa cells. All doses of LH tested had stimulatory effect on estradiol-17β while progesterone secretion by granulosa cells was unaffected.