GDF-9 and bFGF enhance the effect of FSH on the survival, activation, and growth of cattle primordial follicles

This study aims to investigate the effects of follicle stimulating hormone (FSH) in combination with growth and differentiation factor-9 (GDF-9) or basic fibroblast growth factor (bFGF) on the activation, survival and growth of cattle primordial follicles. Ovarian tissues were cultured for 3, 7, 14, 22 days in α minimum essential medium (α-MEM) supplemented with FSH, FSH+GDF-9 or FSH+bFGF. Non-cultured and cultured ovarian fragments were processed for histological and TUNEL analysis. Compared to the FSH medium, the results showed FSH+GDF-9 medium increased the percentage of primary follicles in all culture periods and secondary follicles after 14 days of culture (P<0.05), meanwhile the diameter of primary and secondary follicles were also observed to increase in this medium after 7 days of cultures (P<0.05). FSH+bFGF medium appeared to increase the percentage of primary follicles after 14 days of culture and secondary follicles at day 14 of culture than FSH medium (P<0.05). Furthermore, the FSH+GDF-9 and FSH+bFGF mediums had a greater percentage of normal follicles, and lesser apoptotic cell rates than FSH medium. The results first indicated that FSH in combination with GDF-9 or bFGF can improve the survival, activation, and growth of cattle primordial follicles after the long-term culture of ovarian cortex.