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2014 ; 15
(5
): 7139-57
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Generation of bladder urothelium from human pluripotent stem cells under
chemically defined serum- and feeder-free system
#MMPMID24776760
Kang M
; Kim HH
; Han YM
Int J Mol Sci
2014[Apr]; 15
(5
): 7139-57
PMID24776760
show ga
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.