Derivation of neural crest cells from human pluripotent stem cells Journal Article
| Authors: | Lee, G.; Chambers, S. M.; Tomishima, M. J.; Studer, L. |
| Article Title: | Derivation of neural crest cells from human pluripotent stem cells |
| 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. |
| Keywords: | genetics; flow cytometry; methodology; cell proliferation; cytology; metabolism; biological model; embryonic stem cell; cell differentiation; in vitro study; validation study; bacterial artificial chromosome; chromosomes, artificial, bacterial; cell culture techniques; embryonic stem cells; neural crest; pluripotent stem cell; pluripotent stem cells; schwann cell; culture technique; cell separation; coculture; coculture techniques; schwann cells; models, neurological |
| Journal Title: | Nature Protocols |
| Volume: | 5 |
| Issue: | 4 |
| ISSN: | 1754-2189 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2010-01-01 |
| Start Page: | 688 |
| End Page: | 701 |
| Language: | English |
| DOI: | 10.1038/nprot.2010.35 |
| PUBMED: | 20360764 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 3" - "Export Date: 20 April 2011" - "Source: Scopus" |
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