Derivation of neural crest cells from human pluripotent stem cells

Gabsang Lee, Stuart M. Chambers, Mark J. Tomishima, Lorenz Studer

Research output: Contribution to journalArticle

Abstract

Human pluripotent stem cell (hPSC)-derived neural crest (NC) cells present a valuable tool for modeling aspects of human NC development, including cell fate specification, multipotency and cell migration. hpsc-derived Nc cells are also suitable for modeling human disease and as a renewable cell source for applications in regenerative medicine. Here we provide protocols for the step-wise differentiation of human embryonic stem cells (hEscs) or human induced pluripotent stem cells (hipscs) into neuroectodermal and Nc cells using either the Ms5 coculture system or a novel defined culture method based on pharmacological inhibition of bone morphogenetic protein and transforming growth factor-β signaling pathways. Furthermore, we present protocols for the purification and propagation of hPSc-Nc cells using flow cytometry and defined in vitro culture conditions. our protocol has been validated in multiple independent hESc and hipsc lines. The average time required for generating purified hPSc-Nc precursors using this protocol is 2-5 weeks.

Original languageEnglish (US)
Pages (from-to)688-701
Number of pages14
JournalNature protocols
Volume5
Issue number4
DOIs
StatePublished - Apr 2010
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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