Isolation and propagation of enteric neural crest progenitor cells from mouse embryonic stem cells and embryos

Item Type:	Article
Title:	Isolation and propagation of enteric neural crest progenitor cells from mouse embryonic stem cells and embryos
Creators Name:	Kawaguchi, J. and Nichols, J. and Gierl, M.S. and Faial, T. and Smith, A.
Abstract:	Neural crest is a source of diverse cell types, including the peripheral nervous system. The transcription factor Sox10 is expressed throughout early neural crest. We exploited Sox10 reporter and selection markers created by homologous recombination to investigate the generation, maintenance and expansion of neural crest progenitors. Sox10-GFP-positive cells are produced transiently from mouse embryonic stem (ES) cells by treatment with retinoic acid in combination with Fgf8b and the cytokine leukaemia inhibitory factor (Lif). We found that expression of Sox10 can be maintained using noggin, Wnt3a, Lif and endothelin (NWLE). ES cell-derived Sox10-GFP-positive cells cultured in NWLE exhibit molecular markers of neural crest progenitors. They differentiate into peripheral neurons in vitro and are able to colonise the enteric network in organotypic gut cultures. Neural crest cells purified from embryos using the Sox10 reporter also survive in NWLE, but progressively succumb to differentiation. We therefore applied selection to eliminate differentiating cells. Sox10-selected cells could be clonally expanded, cryopreserved, and multiplied for over 50 days in adherent culture. They remained neurogenic in vitro and in foetal gut grafts. Generation of neural crest from mouse ES cells opens a new route to the identification and validation of determination factors. Furthermore, the ability to propagate undifferentiated progenitors creates an opportunity for experimental dissection of the stimuli and molecular circu that govern neural crest lineage progression. Finally, the demonstration of robust enteric neurogenesis provides a system for investigating and modelling cell therapeutic approaches to neurocristopathies such as Hirschsprung's disease.
Keywords:	Embryonic Stem Cells, Enteric Nervous System, Neural Crest, Animals, Mice
Source:	Development
ISSN:	0950-1991
Publisher:	Company of Biologists
Volume:	137
Number:	5
Page Range:	693-704
Date:	March 2010
Official Publication:	https://doi.org/10.1242/dev.046896
PubMed:	View item in PubMed

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