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F3369 Standard Guide for Assessing the Skeletal Myoblast Phenotype

2019

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  • Título:
    F3369 Standard Guide for Assessing the Skeletal Myoblast Phenotype
  • Descrição: 5.1 This guide describes markers involved in myoblast differentiation that can be used to screen stem cells to help define myogenic capacity. Stem cells include pluripotent and multipotent stem cells capable of differentiating into several different mesenchymal cells, including skeletal muscle myoblasts. 5.2 To assess myogenesis in cells derived and not derived from muscle, markers are measured to accurately define the changes in transcription and structural proteins that regulate differentiation, fusion, and myotube formation. Discussion of these markers is important to understand why they are recommended. 5.3 Myogenic Differentiation:  5.3.1 Myogenic differentiation is a highly regulated process controlled by paired box (Pax) transcription factors and the myogenic regulatory factor (MRF) family. During early differentiation in adults, myogenic progenitors such as activated satellite cells or myoblasts express Pax3 and Pax7. Pax3 and Pax7 transcription factors switch the cells toward a myogenic fate, and repress myocyte differentiation (2), priming the cell for later MRFs. To form muscle, the family of MRFs is required to terminally differentiate myoblasts and form myofibers. These regulatory proteins belong to a superfamily of basic helix-loop-helix transcription factors that consists of myogenic differentiation factor 1 (Myod1), myogenic factor 5 (Myf5), myogenin (Myog), and myogenic factor 6 (Myf6). In the initial stages of myogenic differentiation, Myod1 and Myf5 are the first MRFs to be expressed, and trigger increased production of Myog and Myf6 (3). Increased intracellular Myog and Myf6 induces terminal differentiation of myoblasts into myocytes, leading to fused myotubes. 5.4 Forming Myotubes:  5.4.1 While myogenic markers describe differentiation, fusion into multinucleated myotubes is an important factor in muscle biology. Myoblasts differentiate into a fusogenic phenotype characterized by multiple fusion markers. One marker of note is m-cadherin. M-cadherin is reported to be involved in myoblast fusion and to regulate myotube development (4). Therefore, assessment of fusion markers in addition to myogenic differentiation markers would favor a cell phenotype capable of forming muscle. In support of this, studies have shown that despite expression of myogenic differentiation genes, cells not expressing m-cadherin were unable to fuse and form muscle. These results suggest that in addition to myogenic differentiation markers, fusion markers should be considered given their importance as indicators of whether a cell is able to fuse (5). This guide will enumerate published methods to measure and quantify myoblast fusion markers.
  • Data de criação/publicação: 2019
  • Idioma: Inglês
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  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

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