Development of the conditionally immortalized testicular Sertoli cell line TTE3 expressing Sertoli cell specific genes from mice transgenic for temperature sensitive simian virus 40 large T antigen gene.
OBJECTIVE
We developed and characterized a conditionally immortalized testicular Sertoli cell line from transgenic mice bearing the temperature sensitive simian virus 40 large T antigen gene pSVtsA58.


METHODS
Established cells from 8-week-old male transgenic mice were cultured at a permissive (33C) or nonpermissive (39C) temperature on a collagen type I pre-coated culture vessel. The expression of Sertoli cell specific proteins was analyzed by reverse transcriptase-polymerase chain reaction, immunocytochemical testing and Western blot analysis.


RESULTS
The Sertoli cell line TTE3 grew at 33C but not at 39C. Large T antigen was expressed only in the nuclei at 33C, indicating that the temperature sensitive growth phenotype of the cells arose as a result of the function of temperature sensitive simian virus 40 large T antigen. The cells did not show any colony forming activity in soft agar or form tumors in subcutaneous tissue in nude mice, showing that TTE3 cells were not transformed. The cells expressed messenger RNAs encoding steel factor, inhibin-alpha, transferrin, follicle-stimulating hormone receptor and sulfated glycoprotein-2. Moreover, expression of vimentin and zonula occludens-1 was observed in the cytoplasm and on the boundaries of the cells, respectively. Interestingly expression levels of transferrin and zonula occludens-1 were significantly elevated at 39C.


CONCLUSIONS
TTE3 cells with these unique characteristics should serve as a useful model to study the regulation of Sertoli cell function.
Tabuchi
Yoshiaki
Y
Molecular Genetics Research Center, Department of Urology, Faculty of Medicine, Toyama Medical and Pharmaceutical University, Toyama, Japan.
Ohta
Shoichiro
S
Yanai
Nobuaki
N
Obinata
Masuo
M
Kondo
Takashi
T
Fuse
Hideki
H
Asano
Shinji
S
D000818:Animals; D000957:Antigens, Viral, Tumor / Q000276:immunology*; D015153:Blotting, Western; D002460:Cell Line; D017077:Culture Media, Conditioned; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D010750:Phosphoproteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012708:Sertoli Cells / Q000276:immunology*; D013539:Simian virus 40 / Q000276:immunology*; D013696:Temperature; D013737:Testis / Q000166:cytology*; D014168:Transferrin / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism; D062826:Zonula Occludens-1 Protein
D000957:Antigens, Viral, Tumor; D017077:Culture Media, Conditioned; D008565:Membrane Proteins; D010750:Phosphoproteins; C569043:Tjp1 protein, mouse; D014168:Transferrin; D014746:Vimentin; D062826:Zonula Occludens-1 Protein