DEVELOPMENT AND CHARACTERIZATION OF A PREPUBERTAL RAT SERTOLI-CELL LINE, 93RS2

Sertoli cells in the seminiferous epithelium provide both structural a nd nutritional support to germ cells during spermatogenesis. Primary S ertoli cells in culture are an effective tool for the in vitro study o f Sertoli cell function; however, primary cultures are inherently vari able, time consuming to prepare, expensive, and wasteful of animals. W e therefore developed a Sertoli cell line, called 93RS2, by immortaliz ing primary Sertoli cells derived from prepubertal rats with SV40 tsA2 55. This cell line proliferates at the permissive temperature (32 degr ees C) and has enhanced expression of a differentiated Sertoli cell ph enotype at the nonpermissive temperature (40-41 degrees C). Cytogeneti c analysis demonstrated that 93RS2 has 42 chromosomes per cell, the sa me as a normal rat. mRNA analysis showed that this cell line, when cul tured at a nonpermissive temperature, exhibited increased expression o f transferrin in the presence of testosterone and enhanced expression of sulfated glycoprotein-2. A tumorigenicity assay showed that 93RS2 c ells were temperature-dependent for growth in soft agar and were capab le of forming tumors in nude mice. In conclusion, this rat 93RS2 cell line should be useful for the study of Sertoli cell function.