title: Derivation and expansion of PAX7-positive muscle progenitors from human and mouse embryonic stem cells.
abstract: Cell therapies treating pathological muscle atrophy or damage requires an adequate quantity of muscle progenitor cells (MPCs) not currently attainable from adult donors. Here, we generate cultures of approximately 90% skeletal myogenic cells by treating human embryonic stem cells (ESCs) with the GSK3 inhibitor CHIR99021 followed by FGF2 and N2 supplements. Gene expression analysis identified progressive expression of mesoderm, somite, dermomyotome, and myotome markers, following patterns of embryonic myogenesis. CHIR99021 enhanced transcript levels of the pan-mesoderm gene T and paraxial-mesoderm genes MSGN1 and TBX6; immunofluorescence confirmed that 91% ± 6% of cells expressed T immediately following treatment. By 7 weeks, 47% ± 3% of cells were MYH(+ve) myocytes/myotubes surrounded by a 43% ± 4% population of PAX7(+ve) MPCs, indicating 90% of cells had achieved myogenic identity without any cell sorting. Treatment of mouse ESCs with these factors resulted in similar enhancements of myogenesis. These studies establish a foundation for serum-free and chemically defined monolayer skeletal myogenesis of ESCs.
authors.affiliation: Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
authors.forename: Michael
authors.initials: M
authors.lastname: Shelton
authors.affiliation: Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
authors.forename: Jeff
authors.initials: J
authors.lastname: Metz
authors.affiliation: Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.affiliation: Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON K1H 8L6, Canada; Faculty of Graduate and Postdoctoral Studies, University of Ottawa, Ottawa, ON K1N 6N5, Canada.
authors.forename: Richard L
authors.initials: RL
authors.lastname: Carpenedo
authors.affiliation: Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON K1H 8L6, Canada; Faculty of Graduate and Postdoctoral Studies, University of Ottawa, Ottawa, ON K1N 6N5, Canada.
authors.forename: Simon-Pierre
authors.initials: SP
authors.lastname: Demers
authors.affiliation: Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada; Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON K1H 8L6, Canada; Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON K1H 8L6, Canada; Faculty of Graduate and Postdoctoral Studies, University of Ottawa, Ottawa, ON K1N 6N5, Canada. Electronic address: wstanford@ohri.ca.
authors.forename: William L
authors.initials: WL
authors.lastname: Stanford
authors.affiliation: Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada. Electronic address: iskerjan@uottawa.ca.
authors.forename: Ilona S
authors.initials: IS
authors.lastname: Skerjanc
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors; D006801:Humans; D008648:Mesoderm / Q000166:cytology / Q000187:drug effects / Q000196:embryology; D051379:Mice; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000166:cytology* / Q000378:metabolism; D051763:PAX7 Transcription Factor / Q000032:analysis* / Q000235:genetics; D011725:Pyridines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology
chemical_list: C473711:Chir 99021; D051763:PAX7 Transcription Factor; D011725:Pyridines; D011743:Pyrimidines; D016222:Fibroblast Growth Factor 2; D038362:Glycogen Synthase Kinase 3