In Vitro Maturation of Human Pluripotent Stem Cell-Derived Myotubes

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

Research output: Chapter in Book/Report/Conference proceedingChapter


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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages14
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Funding Information:
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.

Publisher Copyright:
© 2023, Springer Science+Business Media, LLC, part of Springer Nature.


  • Induced pluripotent stem (iPS) cells
  • Myogenic differentiation
  • Myotubes
  • PAX7
  • Small molecules

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural


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