Immortalized human bladder cell line exhibits amiloride-sensitive
sodium absorption.
Perrone, Ronald D., Conrado Johns, Shelley A. Grubman, Elisa Moy, Des
W. Lee, Joseph Alroy, Grannum R. Sant, and Douglas M. Jefferson.
Departments of Medicine, Pathology, Pediatrics and Urology, New
England Medical Center and Tufts University School of Medicine,
Department of Physiology, Tufts University School of Medicine, and
Department of Pathology, Tufts University School of Veterinary
Medicine, Boston, MA 02111
APStracts 2:0130F, 1995.
We have produced a continuous cell line using retroviral transduction
of 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 media 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 - 1000 ohm.cm2, amiloride-sensitive
short-circuit current indicative of active transepithelial sodium
absorption, and functional evidence for basolateral Na,K-ATPase. This
immortalized bladder cell line will facilitate the study of human
bladder epithelial function and the response to diverse physiologic
and pathophysiologic stimuli.
Received 25 April 1995; accepted in final form 17 July 1995.
APS Manuscript Number F135-5.
Article publication pending Am. J. Physiol. (Renal Fluid Electrolyte
Physiology).
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on 10 August 1995.