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Am J Physiol Renal Physiol 270: F148-F153, 1996;
0363-6127/96 $5.00
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AJP - Renal Physiology, Vol 270, Issue 1 148-F153, Copyright © 1996 by American Physiological Society

ARTICLES

Immortalized human bladder cell line exhibits amiloride-sensitive sodium absorption

R. D. Perrone, C. Johns, S. A. Grubman, E. Moy, D. W. Lee, J. Alroy, G. R. Sant and D. M. Jefferson
Department of Medicine, New England Medical Center, Boston, Massachusetts, USA.

We have produced a continuous cell line using retroviral transduction of simian virus 40 (SV40) large T antigen into epithelial cells grown from a cystoscopic bladder biopsy from a female patient with interstitial cystitis. Immortalized urothelial cells are grown in a hormonally supplemented medium in the presence of lethally irradiated NIH-3T3 fibroblast coculture. They maintain their epithelial appearance and are positive for cytokeratins. SV40 large T antigen is localized to the cell nucleus. When grown on Anocell permeable supports, the cells form a complex epithelium with scalloped luminal membranes, apical junctional complexes containing tight junctions, stratification, transepithelial resistance of 500-1,000 omega.cm2, amiloride-sensitive short-circuit current indicative of active transepithelial Na+ absorption, and functional evidence for basolateral Na-K-adenosinetriphosphatase. This immortalized bladder cell line will facilitate the study of human bladder epithelial function and the response to diverse physiological and pathophysiological stimuli.


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