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In Vitro Maturation of Human Pluripotent Stem Cell-Derived Myotubes

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Skeletal Muscle Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2640))

Abstract

Pluripotent stem cells have a multitude of potential applications in the areas of disease modeling, drug screening, and cell-based therapies for genetic diseases, including muscular dystrophies. The advent of induced pluripotent stem cell technology allows for the facile derivation of disease-specific pluripotent stem cells for any given patient. Targeted in vitro differentiation of pluripotent stem cells into the muscle lineage is a key step to enable all these applications. Transgene-based differentiation using conditional expression of the transcription factor PAX7 leads to the efficient derivation of an expandable and homogeneous population of myogenic progenitors suitable for both in vitro and in vivo applications. Here, we describe an optimized protocol for the derivation and expansion of myogenic progenitors from pluripotent stem cells using conditional expression of PAX7. Importantly, we further describe an optimized procedure for the terminal differentiation of myogenic progenitors into more mature myotubes, which are better suited for in vitro disease modeling and drug screening studies.

Ricardo Mondragon-Gonzalez and Sridhar Selvaraj contributed equally to this work.

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Acknowledgments

This work was supported by NIH grants R01 AR071439 and AR055299 (R.C.R.P.). We thank James Kiley for helpful commentaries on this protocol.

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Authors and Affiliations

  1. Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, MN, USA

    Ricardo Mondragon-Gonzalez, Sridhar Selvaraj & Rita C. R. Perlingeiro

  2. Stem Cell Institute, University of Minnesota, Minneapolis, MN, USA

    Rita C. R. Perlingeiro

Authors
  1. Ricardo Mondragon-Gonzalez
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  2. Sridhar Selvaraj
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  3. Rita C. R. Perlingeiro
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Corresponding author

Correspondence to Rita C. R. Perlingeiro .

Editor information

Editors and Affiliations

  1. Stem Cell Institute, University of Minnesota Medical School, Minneapolis, MN, USA

    Atsushi Asakura

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Mondragon-Gonzalez, R., Selvaraj, S., Perlingeiro, R.C.R. (2023). In Vitro Maturation of Human Pluripotent Stem Cell-Derived Myotubes. In: Asakura, A. (eds) Skeletal Muscle Stem Cells. Methods in Molecular Biology, vol 2640. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3036-5_10

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  • DOI: https://doi.org/10.1007/978-1-0716-3036-5_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3035-8

  • Online ISBN: 978-1-0716-3036-5

  • eBook Packages: Springer Protocols

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