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Human Pluripotent Stem Cell Differentiation to Microglia

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Induced Pluripotent Stem (iPS) Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2454))

Abstract

Microglia, the immune cells of the central nervous system (CNS), play critical roles in CNS homeostasis and disease. Mounting evidence has linked aberrant microglial functions to neurodevelopment, neuroinflammatory and neurodegenerative diseases, underlining the need for novel models to investigate human microglia biology. Here we describe a protocol for generating in vitro patient-specific microglia progenitors and microglia-like cells from induced pluripotent stem cells (iPSCs). Our protocol generates microglia progenitor cells in approximately 35 days, which then can further mature into microglia-like cells within two additional weeks. Microglia differentiation is driven by specific growth factors and cytokines in serum-free conditions, resulting in mesodermal progenitors that grow in a monolayer which releases free-floating microglia progenitors. Isolated progenitors can be used in co-culture systems with other neuronal cells, xenotransplanted to generate chimeric mouse models, or further differentiated into adherent microglia-like cells for functional studies.

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Acknowledgments

This work was supported by the New York Stem Cell Foundation Research Institute and by the National Institute on Aging (NIA) grant RF1AG057440.

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Authors and Affiliations

  1. The New York Stem Cell Foundation Research Institute, New York, NY, USA

    Laraib Ijaz, Madhura Nijsure & Valentina Fossati

Authors
  1. Laraib Ijaz
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  2. Madhura Nijsure
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  3. Valentina Fossati
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Corresponding author

Correspondence to Valentina Fossati .

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Editors and Affiliations

  1. Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, ON, Canada

    Andras Nagy

  2. Ottawa, ON, Canada

    Kursad Turksen

1 Electronic Supplementary Materials

Time-lapse video of mature microglia in phase contrast (20×) over a span of 4 h and 30 min capturing microglia motile processes, scanning behavior, and interaction with nearby microglia (MP4 11661 kb)

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Ijaz, L., Nijsure, M., Fossati, V. (2021). Human Pluripotent Stem Cell Differentiation to Microglia. In: Nagy, A., Turksen, K. (eds) Induced Pluripotent Stem (iPS) Cells. Methods in Molecular Biology, vol 2454. Humana, New York, NY. https://doi.org/10.1007/7651_2021_359

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  • DOI: https://doi.org/10.1007/7651_2021_359

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2118-9

  • Online ISBN: 978-1-0716-2119-6

  • eBook Packages: Springer Protocols

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