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1 Department of Biology, University of York, York, United Kingdom; Pyrah Department of Urology, St. James's University Hospital, Leeds, United Kingdom
2 Pyrah Department of Urology, St. James's University Hospital, Leeds, United Kingdom
3 Jack Birch Unit of Molecular Carcinogenesis, University of York, York, United Kingdom
4 Department of Biology, University of York, York, United Kingdom
* To whom correspondence should be addressed. E-mail: js35{at}york.ac.uk.
The urinary bladder and associated tract is lined by urothelium. Once
considered as just an impermeable epithelium, it is becoming evident that urothelium
not only functions as a volume-accommodating urinary barrier but has additional
roles, including sensory signalling. Lack of access to normal human urothelium has
hampered physiological investigation and although cell culture systems have been
developed, there has been a failure to demonstrate that normal human urothelial
(NHU) cells grown in vitro retain the capacity to form a functional differentiated
urothelium. The aim of this study was to develop a biomimetic human urothelium
from NHU cell cultures. Urothelial cells isolated from normal human urothelium and
serially-propagated as monolayers in serum-free culture were homogeneous and
adopted a proliferative, non-differentiated phenotype. In the presence of serum and
physiological concentrations of calcium, these cells could be reproducibly induced to
form stratified urothelia consisting of basal, intermediate and superficial cells, with
differential expression of cytokeratins and superficial tight junctions. Functionally, the
neo-tissues showed characteristics of native urothelium, including high transepithelial
electrical resistance of >3000 
.cm2, apical membrane-restricted amiloride-sensitive
sodium ion channels, basal expression of Na+, K+-ATPase and low diffusive
permeability to urea, water and dextran. This model represents major progress in
developing a biomimetic human urothelial culture model to explore molecular and
functional relationships in normal and dysfunctional bladder physiology.
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