title: Generation of myospheres from hESCs by epigenetic reprogramming.
abstract: 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.
authors.affiliation: Muscle Development and Regeneration Program, Sanford-Burnham Institute for Medical Research; salbini@sanfordburnham.org.
authors.forename: Sonia
authors.initials: S
authors.lastname: Albini
authors.affiliation: Muscle Development and Regeneration Program, Sanford-Burnham Institute for Medical Research; IRCCS Fondazione Santa Lucia; lpuri@sanfordburnham.org.
authors.forename: Pier Lorenzo
authors.initials: PL
authors.lastname: Puri
mesh_terms: D057005:Bioengineering / Q000379:methods*; D002868:Chromosomal Proteins, Non-Histone; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D057890:Epigenomics; D006801:Humans; D018482:Muscle, Skeletal / Q000166:cytology / Q000502:physiology*; D017570:MyoD Protein / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; C491649:SMARCD3 protein, human; D014157:Transcription Factors