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Updated: Aug 20, 2026
Isolation of Conditionally Immortalized Mouse Glomerular Endothelial Cells with Fluorescent Mitochondria
Published on: September 13, 2022
Establishment of immortalized human glomerular endothelial cell lines and their application
Takashi Harada1, Stephen Batsford, Tetsuo Morioka
1Department of Cellular Physiology, Institute of Nephrology, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
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.
