title: Immortalized human bladder cell line exhibits amiloride-sensitive sodium absorption.
abstract: 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.
authors.affiliation: Department of Medicine, New England Medical Center, Boston, Massachusetts, USA.
authors.forename: R D
authors.initials: RD
authors.lastname: Perrone
authors.forename: C
authors.initials: C
authors.lastname: Johns
authors.forename: S A
authors.initials: SA
authors.lastname: Grubman
authors.forename: E
authors.initials: E
authors.lastname: Moy
authors.forename: D W
authors.initials: DW
authors.lastname: Lee
authors.forename: J
authors.initials: J
authors.lastname: Alroy
authors.forename: G R
authors.initials: GR
authors.lastname: Sant
authors.forename: D M
authors.initials: DM
authors.lastname: Jefferson
mesh_terms: D000042:Absorption / Q000187:drug effects; D000584:Amiloride / Q000494:pharmacology*; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D002461:Cell Line, Transformed; D004553:Electric Conductivity; D005260:Female; D006801:Humans; D008854:Microscopy, Electron; D012190:Retroviridae / Q000235:genetics; D012964:Sodium / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000502:physiology; D014161:Transduction, Genetic; D001743:Urinary Bladder / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*
chemical_list: D000952:Antigens, Polyomavirus Transforming; D000584:Amiloride; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase