Establishment of immortalized human glomerular endothelial cell lines and their application.
BACKGROUND
The experimental use of cultured endothelial cells derived from the microvasculature such as glomerular endothelial cells possesses many problems, including limited growth rates, heterogeneity and loss of specific cell properties dependent on culture passage. In this study, we attempted to establish immortalized, human glomerular endothelial cell (HGEC) lines.


METHODS
HGECs of up to 5 passages were transformed by infection with simian virus (SV)-40. After 4-6 weeks the surviving, foci-forming cells were harvested and cloned. Each cell line obtained was examined by immunofluorescence with antibodies to antigens specific for vascular endothelial cells. The expression of adhesion molecules on cells incubated with or without TNF-alpha was also examined by cellular ELISA.


RESULTS
Three of twelve cell lines obtained expressed SV40 large T-antigen and von Willebrand's factor, as well as endothelial cell adhesion molecules including ICAM-1 (CD54), PECAM-1 (CD31) and E-selectin (CD62E). In these cells, ICAM-1 and E-selectin expression was up-regulated by TNF-alpha, as in native cultured HGEC.


CONCLUSIONS
These cell lines maintain the morphologic and functional characteristics of HGEC even after 60 passages. Immortalized HGEC will be useful for research on glomerular cell biology and provide a standardized substrate for anti-endothelial cell antibody detection.
Harada
Takashi
T
Department of Cellular Physiology, Institute of Nephrology, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
Batsford
Stephen
S
Morioka
Tetsuo
T
Yao
Jian
J
Arakawa
Masaaki
M
Gejyo
Fumitake
F
Oite
Takashi
T
D000941:Antigens / Q000032:analysis / Q000096:biosynthesis; D015815:Cell Adhesion Molecules / Q000032:analysis / Q000096:biosynthesis; D002460:Cell Line; D002470:Cell Survival; D042783:Endothelial Cells / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007678:Kidney Glomerulus / Q000166:cytology*; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection; D014170:Transformation, Genetic
D000941:Antigens; D015815:Cell Adhesion Molecules