Myogenic Differentiation of ASCs Using Biochemical and Biophysical Induction

Methods Mol Biol. 2018:1773:123-135. doi: 10.1007/978-1-4939-7799-4_10.

Abstract

Adipose-derived stem/stromal cells (ASCs) constitute a very promising source for cell therapy and tissue engineering approaches as they can be easily obtained in large quantities with comparatively minimal patient discomfort. Moreover, ASCs have multilineage differentiation capacity. Among these, differentiation capacity along the myogenic lineage is of particular interest since myogenic precursors are scarce and obtaining a large number of cells from skeletal muscle biopsies is difficult. Here, we describe a method to effectively induce ASC myogenesis through the combination of biochemical (cocktail including 5-azacytidine and horse serum) and biophysical (dynamic culture via uniaxial cyclic strain) stimulation. This method results in multinucleated cells that are positive in myogenic markers including Pax 3/7, desmin, myoD, and myosin heavy chain.

Keywords: 5-azacytidine; Adipose-derived stem/stromal cell; Cyclic strain; Dynamic culture; Myogenesis; Skeletal muscle.

MeSH terms

  • Adipocytes / cytology
  • Adipocytes / drug effects
  • Adipocytes / immunology
  • Adipose Tissue / cytology
  • Adipose Tissue / drug effects
  • Adipose Tissue / immunology
  • Animals
  • Azacitidine / pharmacology
  • Cell Differentiation / drug effects*
  • Cell Differentiation / immunology
  • Horses / blood
  • Humans
  • Immunohistochemistry
  • Mechanotransduction, Cellular / drug effects*
  • Mechanotransduction, Cellular / immunology
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / immunology
  • Muscle Development / drug effects*
  • Muscle Development / immunology
  • Muscle Fibers, Skeletal / cytology*
  • Muscle Fibers, Skeletal / drug effects
  • Muscle Fibers, Skeletal / immunology
  • Primary Cell Culture
  • Serum / immunology
  • Tissue Engineering

Substances

  • Azacitidine