Generation of myospheres from hESCs by epigenetic reprogramming

Sonia Albini, Pier Lorenzo Puri

Research output: Contribution to journalArticlepeer-review

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 muscles7.

Original languageEnglish
Article numbere51243
JournalJournal of visualized experiments : JoVE
Issue number88
DOIs
Publication statusPublished - Jun 21 2014

Keywords

  • Bioengineering
  • Cells
  • Chromatin
  • Embryonic Structures
  • Epinegetics
  • hESC
  • Infection
  • Issue 88
  • Lentivirus
  • Musculoskeletal Diseases
  • Musculoskeletal System
  • Myosphere
  • Skeletal Myogenesis
  • Tissues

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)
  • Medicine(all)

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