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Differentiation of Urothelium from Mouse Embryonic Stem Cells in Chemically Defined Conditions

  • Badwi B. Boumelhem
  • Stuart T. Fraser
  • Stephen J. AssinderEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2029)

Abstract

The urothelium of the bladder and urethra are derived from the definitive endoderm during development. Cellular signaling molecules important to the developmental specification of the urothelium are also implicated in the dysregulation of the tissue repair mechanism characteristic of bladder disease. Hence, a complete understanding of the regulation of urothelium development is central to understanding the processes of bladder disease, and in development of simple chemically defined methods for use in regenerative medicine. Key to this is a suitable in vitro model that readily allows for the prosecution of biologically pertinent questions. Here a method for differentiating urothelium from mouse embryonic stem cells in chemically defined conditions is described. The method includes a description of flow cytometry and RT-PCR analysis of definitive endoderm markers Cxcr4, c-Kit, and FoxA2, and of terminally differentiated urothelial cell markers Upk1b and Upk2.

Key words

Urothelium Uroepithelium Bladder Uroplakin Interstitial cystitis/painful bladder syndrome Cancer 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Badwi B. Boumelhem
    • 1
  • Stuart T. Fraser
    • 1
  • Stephen J. Assinder
    • 1
    Email author
  1. 1.Disciplines of Physiology, Anatomy and Histology, School of Medical Science and Bosch InstituteUniversity of SydneyCamperdownAustralia

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