Proliferation of newt spermatogonia by mammalian FSH via Sertoli cells in vitro

Abstract

Our previous studies showed that mammalian follicle-stimulating hormone (FSH) stimulated the proliferation and differentiation of secondary spermatogonia into primary spermatocytes in organ culture of testes fragments from Cynops pyrrhogaster. To elucidate how FSH stimulates spermatogonial proliferation, we studied the interaction of spermatogonia with somatic cells in the presence or absence of FSH in cultures of aggregated cells derived from fractionated cell populations. When spermatogonia or those with somatic cells were cultured, they formed aggregates with themselves or with somatic cells, respectively. Most of the somatic cells in the aggregates were Sertoli cells, judged by the lipid droplets in their cytoplasm. [3H]thymidine incorporation into aggregates and autoradiography demonstrated spermatogonial proliferation that was enhanced in the presence of FSH and somatic cells (most probably Sertoli cells), but not in the absence of FSH or in the presence of luteinizing hormone (LH). To examine whether direct contact between spermatogonia and Sertoli cells is indispensable for the stimulation of spermatogonial proliferation by Sertoli cells, the two cell types were separated by a 0.4 micron pore filter in two compartments of a bicameral chamber. In this case, Sertoli cells did not stimulate spermatogonial proliferation in the presence of FSH, indicating that direct contact between spermatogonia and Sertoli cells is indispensable for the stimulation of spermatogonial proliferation by Sertoli cells. Finally, Sertoli cells isolated from the testes from the primary spermatocyte stage did not stimulate spermatogonial proliferation in the presence of FSH. This result indicated that the function of Sertoli cells with respect to their stimulation of proliferation was stage-specific.