Urothelial cell expansion and differentiation are improved by exposure to hypoxia.
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.
Génie tissulaire et régénération, centre de recherche FRQS du CHU de Québec, Axe Médecine Régénératrice, Centre LOEX de l'Université Laval, Québec, QC, Canada.
Stéphane
S
Chabaud
Génie tissulaire et régénération, centre de recherche FRQS du CHU de Québec, Axe Médecine Régénératrice, Centre LOEX de l'Université Laval, Québec, QC, Canada.
Ingrid
I
Saba
Génie tissulaire et régénération, centre de recherche FRQS du CHU de Québec, Axe Médecine Régénératrice, Centre LOEX de l'Université Laval, Québec, QC, Canada.|Programme Analyses Biologique et Biochimiques, Institut Universitaire de Technologie de Laval, Laval, France.
Clément
C
Baratange
Génie tissulaire et régénération, centre de recherche FRQS du CHU de Québec, Axe Médecine Régénératrice, Centre LOEX de l'Université Laval, Québec, QC, Canada.|Programme Analyses Biologique et Biochimiques, Institut Universitaire de Technologie de Laval, Laval, France.
Brice
B
Boiroux
Génie tissulaire et régénération, centre de recherche FRQS du CHU de Québec, Axe Médecine Régénératrice, Centre LOEX de l'Université Laval, Québec, QC, Canada.
Maude
M
Leclerc
Génie tissulaire et régénération, centre de recherche FRQS du CHU de Québec, Axe Médecine Régénératrice, Centre LOEX de l'Université Laval, Québec, QC, Canada.
Alexandre
A
Rousseau
Génie tissulaire et régénération, centre de recherche FRQS du CHU de Québec, Axe Médecine Régénératrice, Centre LOEX de l'Université Laval, Québec, QC, Canada.
Sara
S
Bouhout
Génie tissulaire et régénération, centre de recherche FRQS du CHU de Québec, Axe Médecine Régénératrice, Centre LOEX de l'Université Laval, Québec, QC, Canada.|Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, Canada.
Stéphane
S
Bolduc
D000943:Antigens, Differentiation / Q000378:metabolism*; D002454:Cell Differentiation; D015687:Cell Hypoxia; D049109:Cell Proliferation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D019459:Urothelium / Q000166:cytology / Q000378:metabolism*
D000943:Antigens, Differentiation
bladder mucosa substitutes; bladder urothelium; cell expansion; epithelial cells; hypoxia; normoxia; primary cell culture; reduced oxygen