Journal
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 88, Pages -Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/51243
Keywords
Bioengineering; Issue 88; Tissues; Cells; Embryonic Structures; Musculoskeletal System; Musculoskeletal Diseases; hESC; epinegetics; Skeletal Myogenesis; Myosphere; Chromatin; Lentivirus; Infection
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Funding
- National Institute of Health/National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) [R01AR056712, R01AR052779, P30 AR061303]
- MDA
- Sanford Children Health Research award
- European Community [241440]
- CIRM
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Generation of a homogeneous and abundant population of skeletal muscle cells from human embryonic stem cells (hESCs) is a requirement for cell-based therapies and for a disease in a dish model of human neuromuscular diseases. Major hurdles, such as low abundance and heterogeneity of the population of interest, as well as a lack of protocols for the formation of three-dimensional contractile structures, have limited the applications of stem cells for neuromuscular disorders. We have designed a protocol that overcomes these limits by ectopic introduction of defined factors in hESCs - the muscle determination factor MyoD and SWI/SNF chromatin remodeling complex component BAF60C - that are able to reprogram hESCs into skeletal muscle cells. Here we describe the protocol established to generate hESC-derived myoblasts and promote their clustering into tridimensional miniaturized structures (myospheres) that functionally mimic miniaturized skeletal muscles(7).
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