Development of a conditionally immortalized testicular Sertoli cell line RTS3-3 from adult transgenic rats harboring temperature-sensitive simian virus 40 large T-antigen gene.
Transgenic mice and rats harboring temperature-sensitive simian virus 40 (tsSV40) large T-antigen gene are useful for establishing cell lines from tissues. We succeeded in establishing a conditionally immortalized testicular Sertoli cell line, RT3-3, from adult transgenic rats harboring the oncogene. The cells grew at permissive (33 degrees C) and intermediate (37 degrees C) temperatures but not at nonpermissive temperature (39 degrees C). Large T-antigen was expressed at 33 and 37 degrees C, whereas the expression level was gradually decreased at 39 degrees C, suggesting that the temperature-sensitive growth characteristics arise as a result of the function of tsSV40 large T-antigen. The cells showed biochemical features associate with normal Sertoli cells including expressions of mRNAs of sulfated glycoprotein-2 (SGP-2), transferrin (TF) and steel factor. Quantitative polymerase chain reaction revealed that nonpermissive temperature induced increase in the level of SGP-2. Moreover, levels of SGP-2 and/or TF were significantly elevated in the cells treatment with sodium butyrate and retinoic acid, inducers of cellular differentiation. To our knowledge, this is the first report of the establishment of a testicular Sertoli cell line from the transgenic rats. Thus, the conditionally immortalized cell line RTS3-3 with unique characteristics may serve as good experimental in vitro models for basic and applied biology of testicular Sertoli cells.
Tabuchi
Yoshiaki
Y
Life Science Research Center, Toyama Medical and Pharmaceutical University, 2630 Sugitani, Toyama City, Toyama 930-0194, Japan. ytabu@ms.toyama-mpu.ac.jp
Takahashi
Ri-ichi
R
Ueda
Masatsugu
M
Obinata
Masuo
M
D000818:Animals; D030801:Animals, Genetically Modified; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D001831:Body Temperature / Q000502:physiology*; D020148:Butyric Acid / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002455:Cell Division / Q000187:drug effects / Q000235:genetics*; D002461:Cell Line, Transformed / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051152:Clusterin; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics; D008297:Male; D008954:Models, Biological; D018832:Molecular Chaperones / Q000235:genetics; D009857:Oncogenes / Q000235:genetics*; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012708:Sertoli Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D019089:Stem Cell Factor / Q000235:genetics; D014168:Transferrin / Q000235:genetics; D014212:Tretinoin / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
D000952:Antigens, Polyomavirus Transforming; D051152:Clusterin; D006023:Glycoproteins; D018832:Molecular Chaperones; D012333:RNA, Messenger; D019089:Stem Cell Factor; D014168:Transferrin; D020148:Butyric Acid; D014212:Tretinoin