Abstract
Tissue-engineering offers the opportunity to produce hybrid tissues in vitro. The induction of bladder urothelial cells (BUCs) differentiation in vitro has been assessed by several research groups to build bladder models for fundamental studies and clinical applications. However, BUC induction of advanced differentiation in culture remains a challenging task. To reach this goal, optimal culture conditions are required, notably the use of specific additives as well as proper mesenchymal support. The best positive control for BUCs functional state monitoring is native urothelium collected from healthy bladder samples. In order to establish the best culture conditions to maintain and promote BUC differentiated state, native urothelia were cultured on various mesenchymes. Native bladder mesenchymes were used as controls for the maintenance of native urothelia. Histological and ultrastructural analyses showed the necessity to have a cellularized mesenchyme for rapid formation of a pseudostratified urothelium, allowing apical membrane rearrangement of the superficial cells in culture. Taken together, the results strongly suggest that it is possible to conserve the integrity of urothelia in vitro and, thus, potentially use them for eventual clinical applications and pharmacological investigations.
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Acknowledgments
The authors are grateful to Dr. Tung-Tien Sun, Professor of Cell Biology, Biochemistry & Molecular Pharmacology and Urology (Alexandria Center for Life Sciences, New York University Medical School, NY) for providing anti-uroplakin antibody. They also thank Dr. Francine Goulet for very helpful technical tips and critical review of the manuscript, Dr. Richard Janvier for his precious contribution in electron microscopy analyses, the service of anatomopathology of CHU de Québec, for immunolabeling and histology analyses, and Anne-Marie Moisan for providing the porcine urine and bladder specimen. This study was supported by grants from the “Fonds de la Recherche en Santé du Québec” and the Canadian Institutes of Health Research. Dr. Stéphane Bolduc has received grants from the Kidney Foundation and was a recipient of the Canadian Urological Association Scholarship Fund.
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Bouhout, S., Tremblay, J. & Bolduc, S. Maintenance of bladder urothelia integrity and successful urothelialization of various tissue-engineered mesenchymes in vitro. In Vitro Cell.Dev.Biol.-Animal 51, 922–931 (2015). https://doi.org/10.1007/s11626-015-9923-x
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DOI: https://doi.org/10.1007/s11626-015-9923-x