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Nat Protoc. 2010 Apr;5(4):688-701. doi: 10.1038/nprot.2010.35. Epub 2010 Mar 18.

Derivation of neural crest cells from human pluripotent stem cells.

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  • 1Developmental Biology Program, Sloan-Kettering Institute, New York, NY, USA.

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-beta 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.

PMID:
20360764
[PubMed - indexed for MEDLINE]
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