In vitro proliferation of ICR mouse sertoli cells supplemented with recombinant h-FSH (rh-FSH) at different concentrations

Abstract

Objective: Sertoli cells are important nurse cells in the seminiferous tubules for male germ cell growth and differentiation into spermatozoa. In contrast to the germ cells, Sertoli cells have receptors for follicle stimulating hormone, which are the main hormonal regulators of spermatogenesis. Hormonal control of spermatogenesis involves control of the maturation and supporting activities of Sertoli cells. Impairment of Sertoli cell function has severe consequences for spermatogenesis. In adult testes, the final number of Sertoli cells correlates with the increase of germ cell production and total sperm output. In this study, we have cultured Sertoli cells for studying the growth kinetics of these cells in culture condition in vitro. Design: With the aim of setting up a proliferation assay to investigate the control of Sertoli cell division, we used an easy and rapid technique, based on the use of selective lectin binding which allows to obtain functional Sertoli cells. Materials/Methods: Datura stramonium agglutin (DSA)-coated dish were used to rapidly single Sertoli cells from immature mouse testis. Isolated Sertoli cells were cultured for 17 days in Eagle’s minimal essential medium (EMEM) with supplements according treatment groups. The proliferative activity of Sertoli cells to FSH responsiveness was examined at each different concentration( 0, 37.5, 75, 150, 300 and 600 mIU/ml) of recombinant h-FSH (rh-FSH). The cell purity of Sertoli cell cultured was examined by immunocytochemical procedure using mouse monoclonal anticytokeratin Pan. Results: The rh-FSH dose of 37.5 mIU/ml and 75 mIU/ml showed the dose dependent increase of Sertoli cells. The supplemented rh-FSH was found to stimulate Sertoli cell proliferation in culture condition. But, the proliferation of cells in rh-FSH dose of 300 mIU/ml and 600 mIU/ml were not showed in this study. Hyper-dose of rh-FSH was showed detrimental effect of the Sertoli cell proliferation. Conclusions: The Sertoli cell culture using immunocytochemistry and DSA-coated dish is useful tool in studies of growth kinetics and cell proliferation in vitro. Also the optimal dose of rh-FSH was important factor for Sertoli cell proliferation.