Skeletal Muscle Cell Growth Alters the Lipid Composition of Extracellular Vesicles

We sought to characterize the lipid profile of skeletal muscle cell-derived Extracellular Vesicles (EVs) to determine if a hypertrophic stimulus would affect the lipid composition of C2C12 myotube-derived EVs. Analyses included C2C12 murine myoblasts differentiated into myotubes and treated with Ins...

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Bibliographic Details
Published in:Membranes (Basel) 2021-08, Vol.11 (8), p.619
Main Authors: Valentino, Taylor R, Rule, Blake D, Mobley, C Brooks, Nikolova-Karakashian, Mariana, Vechetti, Ivan J
Format: Article
Language:English
Subjects:
Antibiotics
Biomarkers
Biosynthesis
CD63 antigen
CD81 antigen
Cell culture
Ceramide
Composition
Culture media
Diglycerides
Eutrophication
EV biogenesis
Extracellular vesicles
Gene expression
Growth factors
Insulin
Insulin-like growth factor I
Lipid composition
Lipids
Mass spectrometry
Mass spectroscopy
Metabolites
MicroRNAs
muscle cell
muscle growth
Muscles
Musculoskeletal system
Myoblasts
Myotubes
Nanoparticles
Physiology
Plasma
Skeletal muscle
Sphingosine 1-phosphate
Transmission electron microscopy
Vesicles
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Summary:We sought to characterize the lipid profile of skeletal muscle cell-derived Extracellular Vesicles (EVs) to determine if a hypertrophic stimulus would affect the lipid composition of C2C12 myotube-derived EVs. Analyses included C2C12 murine myoblasts differentiated into myotubes and treated with Insulin-Like Growth Factor 1 (IGF-1) for 24 h to induce hypertrophic growth. EVs were isolated from cell culture media, quantified using Nanoparticle Tracking Analysis (NTA) and analyzed using Transmission Electron Microscopy (TEM). EVs were homogenized and lipids extracted for quantification by Mass Spectrometry followed by downstream lipid class enrichment and lipid chain analysis. IGF-1 treatment elicited an increase in CD63 and CD81 levels (39% and 21%) compared to the controls (16%), respectively. Analysis revealed that skeletal muscle-derived EVs are enriched in bioactive lipids that are likely selectively incorporated into EVs during hypertrophic growth. IGF-1 treatment of myotubes had a significant impact on the levels of diacylglycerol (DG) and ceramide (Cer) in secreted EVs. Specifically, the proportion of unsaturated DG was two- to three-fold higher in EVs derived from IGF-treated cells, as compared to those from control cells. The levels of saturated DG were unaffected. Selective increases were similarly seen in C16- and C24-Cer but not in other species. Levels of free sphingoid bases tended to decrease, while those of sphingosine-1-phosphate was unaffected. Our results suggest that the lipid composition and biogenesis of skeletal muscle-derived EVs, are specific and highly selective during hypertrophic growth.
ISSN:2077-0375
2077-0375
DOI:10.3390/membranes11080619