title: Efficient derivation and inducible differentiation of expandable skeletal myogenic cells from human ES and patient-specific iPS cells.
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.
authors.affiliation: Department of Cell and Developmental Biology, University College London, London, UK.
authors.forename: Sara M
authors.initials: SM
authors.lastname: Maffioletti
authors.affiliation: Department of Cell and Developmental Biology, University College London, London, UK.
authors.forename: Mattia F M
authors.identifier: 0000000293317394
authors.initials: MF
authors.lastname: Gerli
authors.affiliation: Department of Cell and Developmental Biology, University College London, London, UK.
authors.forename: Martina
authors.initials: M
authors.lastname: Ragazzi
authors.affiliation: Department of Gene Therapy and Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium.
authors.forename: Sumitava
authors.initials: S
authors.lastname: Dastidar
authors.affiliation: 1] Department of Cell and Developmental Biology, University College London, London, UK. [2] Present address: Institute of Child Health, University College London, London, UK.
authors.forename: Sara
authors.initials: S
authors.lastname: Benedetti
authors.affiliation: 1] Department of Gene Therapy and Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium. [2] Department of Cardiovascular Sciences, Center for Molecular and Vascular Biology, University of Leuven (KU Leuven), Leuven, Belgium.
authors.forename: Mariana
authors.initials: M
authors.lastname: Loperfido
authors.affiliation: 1] Department of Gene Therapy and Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium. [2] Department of Cardiovascular Sciences, Center for Molecular and Vascular Biology, University of Leuven (KU Leuven), Leuven, Belgium.
authors.forename: Thierry
authors.initials: T
authors.lastname: VandenDriessche
authors.affiliation: 1] Department of Gene Therapy and Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium. [2] Department of Cardiovascular Sciences, Center for Molecular and Vascular Biology, University of Leuven (KU Leuven), Leuven, Belgium.
authors.forename: Marinee K
authors.initials: MK
authors.lastname: Chuah
authors.affiliation: Department of Cell and Developmental Biology, University College London, London, UK.
authors.forename: Francesco Saverio
authors.identifier: 0000000153217682
authors.initials: FS
authors.lastname: Tedesco
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D015925:Cryopreservation; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D024510:Muscle Development; D017570:MyoD Protein / Q000235:genetics / Q000378:metabolism; D032448:Myoblasts, Skeletal / Q000166:cytology* / Q000378:metabolism
chemical_list: D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein