Generation of myospheres from hESCs by epigenetic reprogramming

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...

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Bibliographic Details
Published in:Journal of visualized experiments 2014-06 (88), p.e51243-e51243
Main Authors: Albini, Sonia, Puri, Pier Lorenzo
Format: Article
Language:English
Subjects:
Bioengineering
Bioengineering - methods
Embryonic Stem Cells - cytology
Embryonic Stem Cells - physiology
Epigenomics
Humans
Muscle, Skeletal - cytology
Muscle, Skeletal - physiology
MyoD Protein - genetics
Transcription Factors - genetics
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Summary: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.
ISSN:1940-087X
1940-087X
DOI:10.3791/51243