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.
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