Immortalization of rat eustachian tube epithelial cells by adenovirus 12-simian virus 40 hybrid virus.
The eustachian tube epithelial cells play an important role in the initial pathogenesis of otitis media. In order to study the role of the eustachian tube epithelial cells in the pathogenesis of otitis media, we have established a rat eustachian tube epithelial cell line. The cell line was derived by infecting primary cultures of eustachian tube epithelial cells with the adenovirus 12-simian virus 40 (Adl2-SV40) hybrid virus. The immortalized cells have retained the morphological characteristics of the parental cells and show positive staining with anti-cytokeratin antibodies (a marker for epithelial cells), but not with anti-vimentin antibodies (a fibroblast marker). The cells have been in continuous culture for more than 10 months and have undergone 38 passages. Western blotting and cell staining have confirmed the expression of the SV40 T antigen and p53. Chromosomal analysis indicates that the cell line is aneuploid and derived from male rat epithelial cells. Together, our results suggest that the cell line originated from eustachian tube epithelial cells from a male rat and was successfully immortalized by the Ad12-SV40 virus.
Jin
Sunji
S
Laboratory of Immunology, National Institute on Deafness and Other Communication Disorders, Bethesda, Maryland, USA.
Moon
Sung-Kyun
SK
Gu
Xin-Xing
XX
Ueyama
Shigehiro
S
Rhim
Johng S
JS
Lim
David J
DJ
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015153:Blotting, Western; D018929:Cell Culture Techniques; D002460:Cell Line / Q000821:virology; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D003470:Culture Media; D004847:Epithelial Cells; D005064:Eustachian Tube / Q000166:cytology*; D006824:Hybridization, Genetic; D007150:Immunohistochemistry; D007621:Karyotyping; D008297:Male; D010033:Otitis Media / Q000209:etiology; D051381:Rats; D011921:Rats, Inbred WKY; D013539:Simian virus 40 / Q000235:genetics*; D013997:Time Factors
D003470:Culture Media