Immortalization and characterization of a sertoli cell line from the adult rat

Abstract

To facilitate investigations of the regulation of adult Sertoli cell function, we have established a Sertoli cell line from sexually mature Sprague-Dawley rats. The cells were immortalized with the temperature sensitive mutant of the SV40 virus, tsA255. The tsA255 large T antigen is heat labile and efficiently promotes propagation of cells at 33°C (permissive temperature) but is inactive at 40°C (nonpermissive temperature). The established clonal Sertoli cell line (ASC-17D) proliferates indefinitely at the permissive temperature. However, within 48 h at the nonpermissive temperature, cell proliferation ceases. ASC-17D cells show positive staining with antibodies to cytokeratin and vimentin, consistent with the Sertoli cell origin of these cells. Transferrin and sulfated glycoprotein (SGP)-2 mRNAs were nearly undetectable in ASC-17D cells cultured at the permissive temperature, but expression of both mRNAs was induced at the nonpermissive temperature. In contrast, SGP-1 was expressed equally at both the permissive and nonpermissive temperatures. There was no increase in either transferrin or SGP-2 with FSH or dibutyryl cAMP (db-cAMP) treatment at the permissive temperature or with FSH treatment at the nonpermissive temperature. However, the steady-state levels of both of these mRNAs were substantially increased in the presence of db-cAMP at the nonpermissive temperature. In contrast, SGP-1 mRNA was not affected by either FSH or db- cAMP. These results suggest that the ASC-17D cell line is derived from adult Sertoli cells and may be useful for the study of adult Sertoli cell function.