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. 2023 Mar 30;14:1143966. doi: 10.3389/fphys.2023.1143966

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Copyright © 2023 Aas, Øvstebø, Brusletto, Aspelin, Trøseid, Qureshi, Eid, Olstad, Nyman and Haug.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Characterization of myotube derived extracellular vesicles (EVs). Human myotubes were exposed to electrical pulse stimulation (EPS) for 24 h, and cell derived EVs were collected for 24 h thereafter. Size (A) and concentration (C) of exosomes (EXO) and microvesicles (MV) were measured by nanoparticle tracking analysis (NTA). The presence of EV markers on exosomes and MV captured by anti-CD81-coated magnetic beads were detected with PE-conjugated CD81 (B) and CD63 (D) antibodies by flow-cytometry (BD Accuri C6 flow cytometer). Presence of CD9, calnexin, and heat shock protein 70 (Hsc/Hsp70) on exosomes were measured by Western blotting (E). Cell lysates of SW480 cells (SW) were used as control. Transmission electron microscopy (TEM) images of skeletal muscle cell derived EVs (F). 1. Freshly isolated EVs from conditioned media from human myotubes, scale bar 1 µm. 2. Close up of framed EVs in picture 1, scale bar 200 nm. Data are presented as mean ± SEM (n = 6 in each group). MFI = mean fluorescence intensity.