Efficient derivation and inducible differentiation of expandable skeletal myogenic cells from human ES and patient-specific iPS cells

Sara M. Maffioletti, Mattia F M Gerli, Martina Ragazzi, Sumitava Dastidar, Sara Benedetti, Mariana Loperfido, Thierry Vandendriessche, Marinee K. Chuah, Francesco Saverio Tedesco

Research output: Contribution to journalArticle

Abstract

Skeletal muscle is the most abundant human tissue; therefore, an unlimited availability of myogenic cells has applications in regenerative medicine and drug development. Here we detail a protocol to derive myogenic cells from human embryonic stem (ES) and induced pluripotent stem (iPS) cells, and we also provide evidence for its extension to human iPS cells cultured without feeder cells. The procedure, which does not require the generation of embryoid bodies or prospective cell isolation, entails four stages with different culture densities, media and surface coating. Pluripotent stem cells are disaggregated to single cells and then differentiated into expandable cells resembling human mesoangioblasts. Subsequently, transient Myod1 induction efficiently drives myogenic differentiation into multinucleated myotubes. Cells derived from patients with muscular dystrophy and differentiated using this protocol have been genetically corrected, and they were proven to have therapeutic potential in dystrophic mice. Thus, this platform has been demonstrated to be amenable to gene and cell therapy, and it could be extended to muscle tissue engineering and disease modeling.

Original languageEnglish
Pages (from-to)941-958
Number of pages18
JournalNature Protocols
Volume10
Issue number7
DOIs
Publication statusPublished - Jul 27 2015

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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    Maffioletti, S. M., Gerli, M. F. M., Ragazzi, M., Dastidar, S., Benedetti, S., Loperfido, M., Vandendriessche, T., Chuah, M. K., & Tedesco, F. S. (2015). Efficient derivation and inducible differentiation of expandable skeletal myogenic cells from human ES and patient-specific iPS cells. Nature Protocols, 10(7), 941-958. https://doi.org/10.1038/nprot.2015.057