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Directed Differentiation of Neural-stem cells and Subtype-Specific Neurons from hESCs

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Cellular Programming and Reprogramming

Part of the book series: Methods in Molecular Biology ((MIMB,volume 636))

Abstract

We describe a chemically defined protocol for efficient differentiation of human embryonic stem cells (hESCs) to neural epithelial cells and then to functional spinal motor neurons. This protocol comprises four major steps. Human ESCs are differentiated without morphogens into neuroepithelial cells that form neural tube-like rosettes in the first 2 weeks. The neuroepithelial cells are then specified to OLIG2-expressing motoneuron progenitors in the presence of retinoic acid (RA) and sonic hedgehog (SHH) in the following 2 weeks. These OLIG2 progenitors generate postmitotic, HB9 expressing motoneurons at the fifth week and mature to functional motor neurons thereafter. The protein factor SHH can be replaced by a small molecule purmorphamine in the entire process, which may facilitate potential clinical applications. This protocol has been shown equally effective in generating motor neurons from human induced pluropotent stem (iPS) cells.

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Author information

Authors and Affiliations

  1. Department of Anatomy and Department of Neurology, Waisman Center, School of Medicine and Public Health, the WiCell Institute, University of Wisconsin-Madison, Madison, WI, USA

    Bao-Yang Hu & Su-Chun Zhang

Authors
  1. Bao-Yang Hu
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  2. Su-Chun Zhang
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Corresponding author

Correspondence to Su-Chun Zhang .

Editor information

Editors and Affiliations

  1. The Scripps Research Institute, N. Torrey Pines Road 10550, La Jolla, 92037, U.S.A.

    Sheng Ding

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Hu, BY., Zhang, SC. (2010). Directed Differentiation of Neural-stem cells and Subtype-Specific Neurons from hESCs. In: Ding, S. (eds) Cellular Programming and Reprogramming. Methods in Molecular Biology, vol 636. Humana Press. https://doi.org/10.1007/978-1-60761-691-7_8

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  • DOI: https://doi.org/10.1007/978-1-60761-691-7_8

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-690-0

  • Online ISBN: 978-1-60761-691-7

  • eBook Packages: Springer Protocols

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