Isolation and Characterization of Neural Crest Stem Cells Derived From In Vitro–Differentiated Human Embryonic Stem Cells

Jiang, Xiaohua; Gwye, Ynnez; McKeown, Sonja J.; Bronner‐Fraser, Marianne; Lutzko, Carolyn; Lawlor, Elizabeth R.

doi:10.1089/scd.2008.0362

Cited by 130 publications

(125 citation statements)

References 36 publications

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“…Other studies have relied on surface markers such as HNK1 (human natural killer-1) and p75NTR (NGFR) (Fukuta et al, 2014;Jiang et al, 2009;Lee et al, 2010Lee et al, , 2007Menendez et al, 2011;Umeda et al, 2012), that are respectively known to label only few human NC cells or to have robust expression in non-NC cell populations (Betters et al, 2010). We instead relied on PAX7, TFAP2A and SOX10, which have been identified as robust markers co-expressed in a specific fashion in embryonic human NC (Betters et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

WNT/β-catenin signaling mediates human neural crest induction via a pre-neural border intermediate

et al. 2016

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Neural crest (NC) cells arise early in vertebrate development, migrate extensively and contribute to a diverse array of ectodermal and mesenchymal derivatives. Previous models of NC formation suggested derivation from neuralized ectoderm, via meso-ectodermal, or neural-non-neural ectoderm interactions. Recent studies using bird and amphibian embryos suggest an earlier origin of NC, independent of neural and mesodermal tissues. Here, we set out to generate a model in which to decipher signaling and tissue interactions involved in human NC induction. Our novel human embryonic stem cell (ESC)-based model yields high proportions of multipotent NC cells (expressing SOX10, PAX7 and TFAP2A) in 5 days. We demonstrate a crucial role for WNT/β-catenin signaling in launching NC development, while blocking placodal and surface ectoderm fates. We provide evidence of the delicate temporal effects of BMP and FGF signaling, and find that NC development is separable from neural and/ or mesodermal contributions. We further substantiate the notion of a neural-independent origin of NC through PAX6 expression and knockdown studies. Finally, we identify a novel pre-neural border state characterized by early WNT/β-catenin signaling targets that displays distinct responses to BMP and FGF signaling from the traditional neural border genes. In summary, our work provides a fast and efficient protocol for human NC differentiation under signaling constraints similar to those identified in vivo in model organisms, and strengthens a framework for neural crest ontogeny that is separable from neural and mesodermal fates.

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Section: Discussionmentioning

confidence: 99%

WNT/β-catenin signaling mediates human neural crest induction via a pre-neural border intermediate

et al. 2016

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“…Since osteoblasts may be specified from a neural crest or a mesodermal origin, we also investigated the expression pattern of Mesogenin (Msgn1), a gene that controls presomitic mesoderm maturation downstream of T-Bra (Wittler et al, 2007), as well as the paraxial mesoderm associated gene PDGFRa (Orr-Urtreger et al, 1992;Takakura et al, 1997) and the neural crest marker/ mesenchymal stem cell associated marker p75 (Jiang et al, 2009) a few days further into the differentiation (Fig. 4F).…”

Section: Upstream No Effectorsmentioning

confidence: 99%

NO/beta-catenin crosstalk modulates primitive streak formation prior to embryonic stem cell osteogenic differentiation

Ding

Keller

Martinez

et al. 2012

Journal of Cell Science

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SummaryNitric oxide (NO) has been shown to play a crucial role in bone formation in vivo. We sought to determine the temporal effect of NO on murine embryonic stem cells (ESCs) under culture conditions that promote osteogenesis. Expression profiles of NO pathway members and osteoblast-specific markers were analyzed using appropriate assays. We found that NO was supportive of osteogenesis specifically during an early phase of in vitro development (days 3-5). Furthermore, ESCs stably overexpressing the inducible NO synthase showed accelerated and enhanced osteogenesis in vitro and in bone explant cultures. To determine the role of NO in early lineage commitment, a stage in ESC differentiation equivalent to primitive streak formation in vivo, ESCs were transfected with a T-brachyury-GFP reporter. Expression levels of T-brachyury and one of its upstream regulators, b-catenin, the major effector in the canonical Wnt pathway, were responsive to NO levels in differentiating primitive streak-like cells. Our results indicate that NO may be involved in early differentiation through regulation of b-catenin and T-brachyury, controlling the specification of primitive-streak-like cells, which may continue through differentiation to later become osteoblasts.

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“…The knowledge obtained might eventually help us to develop cell-replacement therapies for neurocristopathies such as Hirschsprung's disease (Gershon, 1997;Heanue and Pachnis, 2007). Several reports have described the production of neural crest cells from human ES cells (Jiang et al, 2009;Lee et al, 2007;Pomp et al, 2005;Pomp et al, 2008). Neural crest derivatives, such as melanocytes, peripheral neurons, osteoblasts and chondrocytes, have also been reported from murine ES cells (Kawaguchi et al, 2005;Mizuseki et al, 2003;Motohashi et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2010

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SUMMARYNeural 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 3 (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 circuitry 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.

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Isolation and Characterization of Neural Crest Stem Cells Derived From In Vitro–Differentiated Human Embryonic Stem Cells

Cited by 130 publications

References 36 publications

WNT/β-catenin signaling mediates human neural crest induction via a pre-neural border intermediate

WNT/β-catenin signaling mediates human neural crest induction via a pre-neural border intermediate

NO/beta-catenin crosstalk modulates primitive streak formation prior to embryonic stem cell osteogenic differentiation

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

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