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Urothelial cell expansion and differentiation are improved by exposure to hypoxia
Author(s) -
Chabaud Stéphane,
Saba Ingrid,
Baratange Clément,
Boiroux Brice,
Leclerc Maude,
Rousseau Alexandre,
Bouhout Sara,
Bolduc Stéphane
Publication year - 2017
Publication title -
journal of tissue engineering and regenerative medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.835
H-Index - 72
eISSN - 1932-7005
pISSN - 1932-6254
DOI - 10.1002/term.2212
Subject(s) - urothelium , urothelial cell , hypoxia (environmental) , microbiology and biotechnology , keratin , epithelium , tissue engineering , keratin 14 , cell , chemistry , biology , pathology , anatomy , medicine , biochemistry , transgene , genetics , urinary system , organic chemistry , genetically modified mouse , oxygen , gene
Cells obtained from a patient's biopsy have to be expanded after extraction to produce autologous tissues, but standard cell culture conditions often limit their growth or lifespan and could induce early and inadequate cell differentiation. Moreover, it has previously been reported that the air–liquid interface, that induces maturation of the urothelium, stimulated inadequate differentiation associated with aberrant keratin‐14 expression. The aim of this study was to test the benefits of hypoxia during expansion of urothelial cells and maturation of the bladder epithelium in the context of tissue engineering. Bladder mucosa substitutes were reconstructed using the self‐assembly method with urothelial cells (UCs) expanded in normoxia or hypoxia. Hypoxia improved UCs expansion until passage P7, whereas normoxic conditions limited the use of UCs to passage P4. Maturation of the urothelium was also compared in normoxic vs. hypoxic conditions. Using laminin V, p63, Ki‐67, keratin‐5 and ‐14, Claudin‐4 and zonula occludens protein‐1, we show a better organization of the basal UC layer in hypoxia despite a thinner intermediate layer. Finally, barrier function was assessed by permeation tests. Cell culture in hypoxia allowed the generation of bioengineered urological tissue closer to native bladder characteristics, which represents a promising avenue to circumvent the lack of adequate tissues for reconstructive surgery. Copyright © 2017 John Wiley & Sons, Ltd.