Your browser doesn't support javascript.
loading
EV-mediated promotion of myogenic differentiation is dependent on dose, collection medium, and isolation method.
Hanson, Britt; Vorobieva, Ioulia; Zheng, Wenyi; Conceição, Mariana; Lomonosova, Yulia; Mäger, Imre; Puri, Pier Lorenzo; El Andaloussi, Samir; Wood, Matthew J A; Roberts, Thomas C.
Afiliación
  • Hanson B; Department of Paediatrics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
  • Vorobieva I; Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
  • Zheng W; Department of Paediatrics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
  • Conceição M; Institute of Developmental and Regenerative Medicine, University of Oxford, IMS-Tetsuya Nakamura Building, Old Road Campus, Roosevelt Dr, Headington, Oxford OX3 7TY, UK.
  • Lomonosova Y; Department of Laboratory Medicine, Karolinska Institutet, Huddinge SE-141 86, Sweden.
  • Mäger I; Department of Paediatrics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
  • Puri PL; Institute of Developmental and Regenerative Medicine, University of Oxford, IMS-Tetsuya Nakamura Building, Old Road Campus, Roosevelt Dr, Headington, Oxford OX3 7TY, UK.
  • El Andaloussi S; Department of Paediatrics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
  • Wood MJA; Institute of Developmental and Regenerative Medicine, University of Oxford, IMS-Tetsuya Nakamura Building, Old Road Campus, Roosevelt Dr, Headington, Oxford OX3 7TY, UK.
  • Roberts TC; Department of Paediatrics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
Mol Ther Nucleic Acids ; 33: 511-528, 2023 Sep 12.
Article en En | MEDLINE | ID: mdl-37602275
Extracellular vesicles (EVs) have been implicated in the regulation of myogenic differentiation. C2C12 murine myoblast differentiation was reduced following treatment with GW4869 or heparin (to inhibit exosome biogenesis and EV uptake, respectively). Conversely, treatment with C2C12 myotube-conditioned medium enhanced myogenic differentiation. Ultrafiltration-size exclusion liquid chromatography (UF-SEC) was used to isolate EVs and non-EV extracellular protein in parallel from C2C12 myoblast- and myotube-conditioned medium. UF-SEC-purified EVs promoted myogenic differentiation at low doses (≤2 × 108 particles/mL) and were inhibitory at the highest dose tested (2 × 1011 particles/mL). Conversely, extracellular protein fractions had no effect on myogenic differentiation. While the transfer of muscle-enriched miRNAs (myomiRs) has been proposed to mediate the pro-myogenic effects of EVs, we observed that they are scarce in EVs (e.g., 1 copy of miR-133a-3p per 195 EVs). Furthermore, we observed pro-myogenic effects with undifferentiated myoblast-derived EVs, in which myomiR concentrations are even lower, suggestive of a myomiR-independent mechanism underlying the observed pro-myogenic effects. During these investigations we identified technical factors with profound confounding effects on myogenic differentiation. Specifically, co-purification of insulin (a component of Opti-MEM) in non-EV LC fractions and polymer precipitated EV preparations. These findings provide further evidence that polymer-based precipitation techniques should be avoided in EV research.
Palabras clave
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Tipo de estudio: Prognostic_studies Idioma: En Revista: Mol Ther Nucleic Acids Año: 2023 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Tipo de estudio: Prognostic_studies Idioma: En Revista: Mol Ther Nucleic Acids Año: 2023 Tipo del documento: Article Pais de publicación: Estados Unidos