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J Biol Chem, Vol. 274, Issue 21, 15020-15029, May 21, 1999
From the Despite almost 25 years of effort, the
development of a highly differentiated and functionally equivalent cell
culture model of uroepithelial cells has eluded investigators. We have
developed a primary cell culture model of rabbit uroepithelium that
consists of an underlying cell layer that interacts with a collagen
substratum, an intermediate cell layer, and an upper cell layer of
large (25-100 µm) superficial cells. When examined at the
ultrastructural level, the superficial cells formed junctional
complexes and had an asymmetric unit membrane, a hallmark of terminal
differentiation in bladder umbrella cells. These cultured
"umbrella" cells expressed uroplakins and a 27-kDa uroepithelial
specific antigen that assembled into detergent-resistant asymmetric
unit membrane particles. The cultures had low diffusive permeabilities
for water (2.8 × 10
Primary Uroepithelial Cultures
A MODEL SYSTEM TO ANALYZE UMBRELLA CELL BARRIER FUNCTION
§,§,,
,§, and§
Renal-Electrolyte Division of the Department
of Medicine and Laboratory of Epithelial Cell Biology, the
§ Department of Cell Biology and Physiology, ¶ Division
of Pulmonology, University of Pittsburgh, Pittsburgh, Pennsylvania
15261, the Epithelial Biology Unit, Departments of Dermatology,
Pharmacology, and Urology, New York University Medical School,
New York, New York 10016
4 cm/s) and urea (3.0 × 107 cm/s) and high transepithelial resistance (>8000
cm2) was achieved when 1 mM
CaCl2 was included in the culture medium. The cell cultures
expressed an amiloride-sensitive sodium transport pathway and increases
in apical membrane capacitance were observed when the cultures were
osmotically stretched. The described primary rabbit cell culture model
mimics many of the characteristics of uroepithelium found in
vivo and should serve as a useful tool to explore normal
uroepithelial function as well as dysfunction as a result of disease.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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