Immortalization and characterization of a Sertoli cell line from the adult rat.
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 degrees C (permissive temperature) but is inactive at 40 degrees 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.
Roberts
K P
KP
Department of Urologic Surgery, University of Minnesota Medical School, Minneapolis 55455, USA. roberts@lenti.med.umn.edu
Banerjee
P P
PP
Tindall
J W
JW
Zirkin
B R
BR
D000818:Animals; D017209:Apoptosis; D003994:Bucladesine / Q000494:pharmacology; D002455:Cell Division; D002461:Cell Line, Transformed; D051152:Clusterin; D004247:DNA / Q000378:metabolism; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D006023:Glycoproteins / Q000235:genetics; D006358:Hot Temperature; D007633:Keratins / Q000032:analysis; D008297:Male; D018832:Molecular Chaperones; D009154:Mutation; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012708:Sertoli Cells / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D013539:Simian virus 40; D014168:Transferrin / Q000235:genetics; D014746:Vimentin / Q000032:analysis
D051152:Clusterin; D006023:Glycoproteins; D018832:Molecular Chaperones; D012333:RNA, Messenger; D014168:Transferrin; D014746:Vimentin; D003994:Bucladesine; D007633:Keratins; D005640:Follicle Stimulating Hormone; D004247:DNA