Kyle N Hagemann, Rhys S McColl, Jason A C Lovett, Celia Snyman, Kathryn H Myburgh
{"title":"Myoblast and fibroblast derived small extracellular vesicles differentially affect myoblast migration dynamics.","authors":"Kyle N Hagemann, Rhys S McColl, Jason A C Lovett, Celia Snyman, Kathryn H Myburgh","doi":"10.1007/s10974-025-09705-y","DOIUrl":null,"url":null,"abstract":"<p><p>Muscle injury activates satellite cells and fibroblasts, with extracellular vesicles (EVs) mediating the related intercellular communication. The influence of EVs released by either cell type on recipient cell behaviour is still unclear. This study investigated the uptake and effects of EVs derived from C2C12 myoblasts (myo-EVs) and L929 fibroblasts (fibro-EVs) on proliferating myoblasts. Both cell lines were cultured in media largely depleted of FBS-derived EVs. Myo-EVs and fibro-EVs isolated from conditioned media were characterised using conventional methods. Effects of these EVs on myoblast function were assessed via PKH67-labelled EV uptake, proliferation, scratch closure, leading front migration rate and individual cell trajectories and western blot analysis for MyoD and Myogenin. Myoblasts preferentially internalised myo-EVs at 5 h (myo-EVs: 3.41 ± 1.34 vs fibro-EVs: 1.25 ± 1.13 puncta per cell) and 48 h (myo-EVs 16.55 ± 12.60 vs fibro-EVs 9.67 ± 4.88) (both p < 0.05). Under proliferative EV-depleted conditions, added EVs did not change myoblast proliferation. However, the elevated expression of Myogenin indicating a subtle shift toward differentiation. Myo-EVs increased myoblast migration rate into a scratch, compared to controls (13.77 ± 1.7 vs 11.08 ± 2.23 µm/h, p < 0.01), but had no effect under conditions of FBS EV depletion. On the other hand, fibro-EVs increased the speed of individual cells, but negatively affected leading front migration due to impaired myoblast persistence. These findings highlight the importance of cell-specific EV-mediated communication in muscle regeneration. Further, tissue explants used to generate cell-specific EVs for treatment should be free of contaminating cell types.</p>","PeriodicalId":16422,"journal":{"name":"Journal of Muscle Research and Cell Motility","volume":" ","pages":""},"PeriodicalIF":1.7000,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Muscle Research and Cell Motility","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1007/s10974-025-09705-y","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"CELL BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Muscle injury activates satellite cells and fibroblasts, with extracellular vesicles (EVs) mediating the related intercellular communication. The influence of EVs released by either cell type on recipient cell behaviour is still unclear. This study investigated the uptake and effects of EVs derived from C2C12 myoblasts (myo-EVs) and L929 fibroblasts (fibro-EVs) on proliferating myoblasts. Both cell lines were cultured in media largely depleted of FBS-derived EVs. Myo-EVs and fibro-EVs isolated from conditioned media were characterised using conventional methods. Effects of these EVs on myoblast function were assessed via PKH67-labelled EV uptake, proliferation, scratch closure, leading front migration rate and individual cell trajectories and western blot analysis for MyoD and Myogenin. Myoblasts preferentially internalised myo-EVs at 5 h (myo-EVs: 3.41 ± 1.34 vs fibro-EVs: 1.25 ± 1.13 puncta per cell) and 48 h (myo-EVs 16.55 ± 12.60 vs fibro-EVs 9.67 ± 4.88) (both p < 0.05). Under proliferative EV-depleted conditions, added EVs did not change myoblast proliferation. However, the elevated expression of Myogenin indicating a subtle shift toward differentiation. Myo-EVs increased myoblast migration rate into a scratch, compared to controls (13.77 ± 1.7 vs 11.08 ± 2.23 µm/h, p < 0.01), but had no effect under conditions of FBS EV depletion. On the other hand, fibro-EVs increased the speed of individual cells, but negatively affected leading front migration due to impaired myoblast persistence. These findings highlight the importance of cell-specific EV-mediated communication in muscle regeneration. Further, tissue explants used to generate cell-specific EVs for treatment should be free of contaminating cell types.
期刊介绍：
The Journal of Muscle Research and Cell Motility has as its main aim the publication of original research which bears on either the excitation and contraction of muscle, the analysis of any one of the processes involved therein, the processes underlying contractility and motility of animal and plant cells, the toxicology and pharmacology related to contractility, or the formation, dynamics and turnover of contractile structures in muscle and non-muscle cells. Studies describing the impact of pathogenic mutations in genes encoding components of contractile structures in humans or animals are welcome, provided they offer mechanistic insight into the disease process or the underlying gene function. The policy of the Journal is to encourage any form of novel practical study whatever its specialist interest, as long as it falls within this broad field. Theoretical essays are welcome provided that they are concise and suggest practical ways in which they may be tested. Manuscripts reporting new mutations in known disease genes without validation and mechanistic insight will not be considered. It is the policy of the journal that cells lines, hybridomas and DNA clones should be made available by the developers to any qualified investigator. Submission of a manuscript for publication constitutes an agreement of the authors to abide by this principle.
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