RESUMO
The p resent study was designed to examine the effect of different doses of FSH on the growth , steroidogenesis and DNA synthesis of two different sizes of buffalo ovarian preantral follicles (PFs) during in vitro culture . Buffalo ovaries were collected from a local abattoir and PFs of small (<150 μm) and large (>150 μm) sizes were isolated through the microdissection method and cultured in TCM - 199 supplemented with 10% FB S, 1% ITS and 30 ng/ml EGF (control group) at 38.5 °C in 5% CO 2 with maximum humidity. The addition of three different doses of FSH (0.5, 1.0 and 2 .0 μg/ml) in the control medium served as the treatment groups . The culture medium was replenished every third day and spent culture medium was stored at - 30 °C for steroid hormone analysis. The results showed that the supplementation of FSH in the culture medium stimulated (P < 0.05) follicular growth , steroidogenesis and DNA synthesis compared to the control. The addition of FSH at 1.0 μg/ml in the culture medium stimulated better (P < 0.05) follicular growth, DNA synthesis and steroidogenesis compared to 0.5 μg/ml FSH , whereas a higher dose of FSH (2.0 μg/ml) inhibited follicular growth in both sizes of follicles . In conclusion, the present study demonstrate d that FSH plays a key role during the development of buffalo ovarian preantral follicles , as it stimulated i n vitro survival, growth rate, steroidogenesis and DNA synthesis of buffalo preantral follicle s at an optim al dose of 1.0 μg/ml.(AU)
Assuntos
Animais , DNA/análise , Esteroides/química , Hormônio Foliculoestimulante/biossíntese , Búfalos/classificação , Folículo Ovariano/anatomia & histologiaRESUMO
The p resent study was designed to examine the effect of different doses of FSH on the growth , steroidogenesis and DNA synthesis of two different sizes of buffalo ovarian preantral follicles (PFs) during in vitro culture . Buffalo ovaries were collected from a local abattoir and PFs of small (150 μm) sizes were isolated through the microdissection method and cultured in TCM - 199 supplemented with 10% FB S, 1% ITS and 30 ng/ml EGF (control group) at 38.5 °C in 5% CO 2 with maximum humidity. The addition of three different doses of FSH (0.5, 1.0 and 2 .0 μg/ml) in the control medium served as the treatment groups . The culture medium was replenished every third day and spent culture medium was stored at - 30 °C for steroid hormone analysis. The results showed that the supplementation of FSH in the culture medium stimulated (P < 0.05) follicular growth , steroidogenesis and DNA synthesis compared to the control. The addition of FSH at 1.0 μg/ml in the culture medium stimulated better (P < 0.05) follicular growth, DNA synthesis and steroidogenesis compared to 0.5 μg/ml FSH , whereas a higher dose of FSH (2.0 μg/ml) inhibited follicular growth in both sizes of follicles . In conclusion, the present study demonstrate d that FSH plays a key role during the development of buffalo ovarian preantral follicles , as it stimulated i n vitro survival, growth rate, steroidogenesis and DNA synthesis of buffalo preantral follicle s at an optim al dose of 1.0 μg/ml.