Development and characterization of a prepubertal rat Sertoli cell line, 93RS2

Abstract

Sertoli cells in the seminiferous epithelium provide both structural and nutritional support to germ cells during spermatogenesis. Primary Sertoli cells in culture are an effective tool for the in vitro study of Sertoli cell function; however, primary cultures are inherently variable, time consuming to prepare, expensive, and wasteful of animals. We therefore developed a Sertoli cell line, called 93RS2, by immortalizing primary Sertoli cells derived from prepubertal rats with SV40 tsA255. This cell line proliferates at the permissive temperature (32 degrees C) and has enhanced expression of a differentiated Sertoli cell phenotype at the nonpermissive temperature (40-41 degrees C). Cytogenetic analysis demonstrated that 93RS2 has 42 chromosomes per cell, the same as a normal rat. mRNA analysis showed that this cell line, when cultured at a nonpermissive temperature, exhibited increased expression of transferrin in the presence of testosterone and enhanced expression of sulfated glycoprotein-2. A tumorigenicity assay showed that 93RS2 cells were temperature-dependent for growth in soft agar and were capable of forming tumors in nude mice. In conclusion, this rat 93RS2 cell line should be useful for the study of Sertoli cell function.