Effects of follicle-stimulating hormone and 17β-estradiol on proliferation of chicken embryonic ovarian germ cells in culture

Abstract

The effects of follicle-stimulating hormone (FSH) and 17β-estradiol (E 2) on chicken ovarian germ cell proliferation were evaluated through a germ-somatic cell coculture model. Ovarian cells from the left ovaries of 18-day-old chicken embryos were cultured in serum-free McCoy's 5A medium at 39 °C and challenged with FSH (0.25–1.0 IU/mL) or E 2 (10 −8–10 −5M) alone and in combination for 48 h. The number of germ cells was counted, and the proliferating cells were immunolocalized by a specific antibody against proliferating cell nuclear antigen (PCNA). The labeling index (LI) was determined for germ cells. Results revealed that germ cells could survive and kept proliferating under support of somatic cells. Germ cells were localized by expression of a specific antibody for stem cell factor receptor c-kit. Both FSH (0.25–1.0 IU/mL) and E 2 (10 −7–10 −5 M) alone induced a marked increase in germ cell number ( P<0.05), and PCNA-LI of germ cells was greater in FSH-treated groups (0.25–1.0 IU/mL) and E 2-treated groups (10 −8–10 −5 M), compared with vehicle-treated group ( P<0.05). Furthermore, FSH manifested a synergistic effect with E 2 (10 −6–10 −5 M) in stimulating germ cell proliferation. These results indicate that FSH might interact with estrogen to promote ovarian germ cell proliferation in embryonic chickens near hatching.