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Directed Differentiation of Neurons from Human iPSCs for Modeling Neurological Disorders

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Cerebral Cortex Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2794))

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Abstract

Human-induced pluripotent stem cell (hiPSC) technology has enabled comprehensive human cell-based disease modeling in vitro. Due to limited accessibility of primary human neurons as well as species-specific divergence between human and rodent brain tissues, hiPSC-derived neurons have become a popular tool for studying neuronal biology in a dish. Here, we provide methods for transcription factor-driven directed differentiation of neurons from hiPSCs via a neural progenitor cell (NPC) intermediate. Doxycycline-inducible expression of neuron fate-determining transcription factors neurogenin 2 (NGN2) and achaete-scute homolog 1 (ASCL1) enables rapid and controllable differentiation of human neurons for disease modeling applications. The provided method is also designed to improve the reproducibility of human neuron differentiation by reducing the batch-to-batch variation of NPC differentiation and lentiviral transduction.

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Acknowledgments

The authors would like to thank Louise and Herbert Horvitz, the Christopher Family, the Judy and Bernard Briskin Fund, and the Sidell Kagan Foundation for their generosity and forethought. This work was supported by the National Institute of Aging of the National Institutes of Health R01 AG056305, RF1 AG061794, R01 AG072291, and RF1 AG079307 to Y.S. J.C. is a predoctoral scholar in the Stem Cell Biology and Regenerative Medicine Research Training Program of the California Institute for Regenerative Medicine (CIRM).

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Authors and Affiliations

  1. Department of Neurodegenerative Diseases, Beckman Research Institute of City of Hope, Duarte, CA, USA

    Cheng Wang, Jonas Cerneckis & Yanhong Shi

  2. Irell & Manella Graduate School of Biological Sciences, Beckman Research Institute of City of Hope, Duarte, CA, USA

    Jonas Cerneckis & Yanhong Shi

Authors
  1. Cheng Wang
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  2. Jonas Cerneckis
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  3. Yanhong Shi
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Corresponding author

Correspondence to Yanhong Shi .

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Editors and Affiliations

  1. Institute for Developmental Research, Aichi Developmental Disability Center, Aichi, Japan

    Koh-ichi Nagata

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Wang, C., Cerneckis, J., Shi, Y. (2024). Directed Differentiation of Neurons from Human iPSCs for Modeling Neurological Disorders. In: Nagata, Ki. (eds) Cerebral Cortex Development. Methods in Molecular Biology, vol 2794. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3810-1_12

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  • DOI: https://doi.org/10.1007/978-1-0716-3810-1_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3809-5

  • Online ISBN: 978-1-0716-3810-1

  • eBook Packages: Springer Protocols

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