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Int. J. Mol. Sci. 2014, 15(5), 7139-7157; doi:10.3390/ijms15057139
Article

Generation of Bladder Urothelium from Human Pluripotent Stem Cells under Chemically Defined Serum- and Feeder-Free System

1,2
, 2
 and 1,3,*
Received: 17 February 2014; in revised form: 25 March 2014 / Accepted: 11 April 2014 / Published: 25 April 2014
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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Abstract: Human stem cells are promising sources for bladder regeneration. Among several possible sources, pluripotent stem cells are the most fascinating because they can differentiate into any cell type, and proliferate limitlessly in vitro. Here, we developed a protocol for differentiation of human pluripotent stem cells (hPSCs) into bladder urothelial cells (BUCs) under a chemically defined culture system. We first differentiated hPSCs into definitive endoderm (DE), and further specified DE cells into BUCs by treating retinoic acid under a keratinocyte-specific serum free medium. hPSC-derived DE cells showed significantly expressed DE-specific genes, but did not express mesodermal or ectodermal genes. After DE cells were specified into BUCs, they notably expressed urothelium-specific genes such as UPIb, UPII, UPIIIa, P63 and CK7. Immunocytochemistry showed that BUCs expressed UPII, CK8/18 and P63 as well as tight junction molecules, E-CADHERIN and ZO-1. Additionally, hPSCs-derived BUCs exhibited low permeability in a FITC-dextran permeability assay, indicating BUCs possessed the functional units of barrier on their surfaces. However, BUCs did not express the marker genes of other endodermal lineage cells (intestine and liver) as well as mesodermal or ectodermal lineage cells. In summary, we sequentially differentiated hPSCs into DE and BUCs in a serum- and feeder-free condition. Our differentiation protocol will be useful for producing cells for bladder regeneration and studying normal and pathological development of the human bladder urothelium in vitro.
Keywords: pluripotent stem cells; cell differentiation; definitive endoderm; urinary bladder; urothelium pluripotent stem cells; cell differentiation; definitive endoderm; urinary bladder; urothelium
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Kang, M.; Kim, H.H.; Han, Y.-M. Generation of Bladder Urothelium from Human Pluripotent Stem Cells under Chemically Defined Serum- and Feeder-Free System. Int. J. Mol. Sci. 2014, 15, 7139-7157.

AMA Style

Kang M, Kim HH, Han Y-M. Generation of Bladder Urothelium from Human Pluripotent Stem Cells under Chemically Defined Serum- and Feeder-Free System. International Journal of Molecular Sciences. 2014; 15(5):7139-7157.

Chicago/Turabian Style

Kang, Minyong; Kim, Hyeon H.; Han, Yong-Mahn. 2014. "Generation of Bladder Urothelium from Human Pluripotent Stem Cells under Chemically Defined Serum- and Feeder-Free System." Int. J. Mol. Sci. 15, no. 5: 7139-7157.


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