Effects of FSH and LH on steroid production by buffalo (Bubalus bubalis) granulosa cells cultured in vitro under serum-free conditions.
The objective of this study was to examine the effects of FSH and LH on oestradiol-17β and progesterone production by buffalo granulosa cells cultured under serum-free conditions. Granulosa cells (3 × 10(5) ) from small (≤ 5 mm diameter) follicles were cultured for up to 4 days in 48-well plates coated with 3.3 μg/cm(2) fibronectin in Dulbecco's modified Eagle's medium (DMEM) : nutrient mixture F-12 Ham (1 : 1 ratio) supplemented with 10(-7) m androstenedione, 5 μg/ml human apo-transferrin and 0.1% bovine serum albumin, in the presence or absence of FSH or LH (0, 1, 2, 4, 8, 16, 32 or 64 ng/ml each). Basal oestradiol-17β production by granulosa cells from small follicles reduced (p < 0.01) from days 1 to 2 of culture and became undetectable by day 3 and basal progesterone production increased (p < 0.05) from day 1 through day 4 of the culture. Although there was no effect of FSH on day 1 of the culture, FSH at 2, 4, 8 and 16 ng/ml increased (p < 0.05) oestradiol-17β production by granulosa cells from small follicles on day 2. Progesterone secretion was increased (p < 0.05) by all doses of FSH on all days of culture. All doses of LH had no effect on oestradiol-17β or progesterone production by granulosa cells from small follicles on any day of the culture. The results of this study demonstrate a serum-free culture system for buffalo granulosa cells and stimulatory effect of FSH but not LH on steroid hormone production by buffalo granulosa cells under these conditions.
Shanmugam
M
M
DCP Division, Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132 001, India. dr_shan@rediffmail.com
Pandita
S
S
Palta
P
P
D000818:Animals; D002020:Buffaloes / Q000378:metabolism*; D018929:Cell Culture Techniques; D016895:Culture Media, Serum-Free; D004958:Estradiol / Q000378:metabolism*; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D006107:Granulosa Cells / Q000187:drug effects* / Q000378:metabolism; D007986:Luteinizing Hormone / Q000494:pharmacology*; D011374:Progesterone / Q000378:metabolism*
D016895:Culture Media, Serum-Free; D011374:Progesterone; D004958:Estradiol; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone