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Efficient myogenic differentiation of human adipose-derived stem cells by the transduction of engineered MyoD protein.

Abstract
Human adipose-derived stem cells (hASCs) have great potential as cell sources for the treatment of muscle disorders. To provide a safe method for the myogenic differentiation of hASCs, we engineered the MyoD protein, a key transcription factor for myogenesis. The engineered MyoD (MyoD-IT) was designed to contain the TAT protein transduction domain for cell penetration and the membrane-disrupting INF7 peptide, which is an improved version of the HA2 peptide derived from influenza. MyoD-IT showed greatly improved nuclear targeting ability through an efficient endosomal escape induced by the pH-sensitive membrane disruption of the INF7 peptide. By applying MyoD-IT to a culture, hASCs were efficiently differentiated into long spindle-shaped myogenic cells expressing myosin heavy chains. Moreover, these cells differentiated by an application of MyoD-IT fused to myotubes with high efficiency through co-culturing with mouse C2C12 myoblasts. Because internalized proteins can be degraded in cells without altering the genome, the myogenic differentiation of hASCs using MyoD-IT would be a safe and clinically applicable method.
AuthorsMin Sun Sung, Ji-Young Mun, Ohsuk Kwon, Ki-Sun Kwon, Doo-Byoung Oh
JournalBiochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 437 Issue 1 Pg. 156-61 (Jul 19 2013) ISSN: 1090-2104 [Electronic] United States
PMID23810391 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCopyright © 2013 Elsevier Inc. All rights reserved.
Chemical References
  • MyoD Protein
  • Peptides
  • Recombinant Fusion Proteins
Topics
  • Adipose Tissue (cytology)
  • Amino Acid Sequence
  • Animals
  • Cell Differentiation
  • Cell Nucleus (metabolism)
  • Coculture Techniques
  • Humans
  • Intracellular Space (metabolism)
  • Mice
  • Molecular Sequence Data
  • Muscle Development
  • Muscle Fibers, Skeletal (metabolism)
  • MyoD Protein (metabolism)
  • Myoblasts (cytology, metabolism)
  • Peptides (chemistry, metabolism)
  • Protein Engineering
  • Protein Transport
  • Recombinant Fusion Proteins (isolation & purification, metabolism)
  • Solubility
  • Stem Cells (cytology, metabolism)
  • Transduction, Genetic

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