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Isolation and directed differentiation of neural crest stem cells derived from human embryonic stem cells

Nature Biotechnology volume 25, pages 1468–1475 (2007)Cite this article

A Corrigendum to this article was published on 01 July 2008

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Abstract

Vertebrate neural crest development depends on pluripotent, migratory precursor cells. Although avian and murine neural crest stem (NCS) cells have been identified, the isolation of human NCS cells has remained elusive. Here we report the derivation of NCS cells from human embryonic stem cells at the neural rosette stage. We show that NCS cells plated at clonal density give rise to multiple neural crest lineages. The human NCS cells can be propagated in vitro and directed toward peripheral nervous system lineages (peripheral neurons, Schwann cells) and mesenchymal lineages (smooth muscle, adipogenic, osteogenic and chondrogenic cells). Transplantation of human NCS cells into the developing chick embryo and adult mouse hosts demonstrates survival, migration and differentiation compatible with neural crest identity. The availability of unlimited numbers of human NCS cells offers new opportunities for studies of neural crest development and for efforts to model and treat neural crest–related disorders.

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Figure 1: Cells expressing neural crest markers are present in cultures of hES cell–derived neural rosettes and can be induced through extrinsic cues.
Figure 2: Molecular characterization of neural crest precursor cells in hES cell–derived neural rosettes.
Figure 3: Identification of neural crest stem cell potential by clonal assay and sphere-forming assay.
Figure 4: Differentiation of hES cell–derived NCS cells toward peripheral nervous system lineages.
Figure 5: Characterization and differentiation of mesenchymal precursor cells derived from hES cell–derived NCS cells.
Figure 6: In vivo transplantation of hES cell–derived NCS cell progeny.
Figure 7: Schematic illustration for the isolation and differentiation of hES cell–derived NCS cell.

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  • 08 July 2008

    In the version of this article initially published, a reference was missing from the first paragraph. The reference (no. 6) has been added and subsequent references renumbered in the HTML and PDF versions of the article.

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Acknowledgements

We would like to thank Z. Dincer and M. Tomishima for technical advice and the Tri-institutional Stem Cell Research Facility at the Memorial Sloan-Kettering Cancer Center (MSKCC) for help in the time-lapse studies. We also would like to thank J. Itskovitz and M. Amit for providing the I-8 cell line, the MSKCC genomics core for performing microarray hybridizations, and members of the Studer, Tabar and Tomishima labs for helpful discussions. This work was supported through the Tri-Institutional Stem Cell Initiative funded by the Starr Foundation.

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Author notes

  1. Hyesoo Kim and Yechiel Elkabetz: These authors contributed equally to this work.

Authors and Affiliations

  1. Developmental Biology Program, Sloan-Kettering Institute, 1275 York Ave., New York, 10021, New York, USA

    Gabsang Lee, Hyesoo Kim, Yechiel Elkabetz & Lorenz Studer

  2. Department of Neurosurgery, Sloan-Kettering Institute, 1275 York Ave., New York, 10021, New York, USA

    George Al Shamy, Georgia Panagiotakos, Viviane Tabar & Lorenz Studer

  3. Division of Neuroscience, Beckman Research Institute of The City of Hope, 1500 E. Duarte Road, Duarte, 91010, California, USA

    Tiziano Barberi

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Contributions

G.L., T.B., V.T. and L.S. designed the study. G.L., V.T. and L.S. analyzed the data and wrote the manuscript. G.L., H.K., Y.E., G.A.S., G.P. and L.S. performed the experiments.

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Correspondence to Lorenz Studer.

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Lee, G., Kim, H., Elkabetz, Y. et al. Isolation and directed differentiation of neural crest stem cells derived from human embryonic stem cells. Nat Biotechnol 25, 1468–1475 (2007). https://doi.org/10.1038/nbt1365

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