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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Small Extracellular Vesicle Release Following Electrical Pulse Stimulation of C2C12 Myotubes: Effects on microRNA Cargo and Myoblast Migration and Differentiation

by
John S. Hingle
1,
Rhys S. McColl
1,
Ivan J. Vechetti
2 and
Kathryn H. Myburgh
1,*
1
Department of Physiological Sciences, Stellenbosch University, Stellenbosch 7602, South Africa
2
Department of Nutrition and Health Sciences, College of Education and Human Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583, USA
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2026, 27(10), 4320; https://doi.org/10.3390/ijms27104320
Submission received: 15 March 2026 / Revised: 5 May 2026 / Accepted: 6 May 2026 / Published: 12 May 2026
(This article belongs to the Special Issue Current Insights into the Role of Exosomes in Intercellular Communication, 2nd Edition)
Versions Notes

Abstract

The skeletal muscle (SkM) secretome has been widely studied since the establishment of its endocrine function. Extracellular vesicles (EVs) are the most recently identified elements of the SkM secretome. These nano-sized lipid-bound vesicles carry molecular cargo and function as a means of intercellular communication. The effect of exercise on SkM EV micro-RNA cargo (miRNAs) remains a challenge to elucidate. Electrical pulse stimulation (EPS) was applied to C2C12 myotubes at high (30 Hz) and low (2 Hz) frequencies. EVs released during 10 h of stimulation were isolated and characterized and used to treat myoblasts. Their miRNA cargo was sequenced. EVs were used to treat myoblasts (2.19 × 108 EVs per mL) to determine the effects on myoblast migration and differentiation. Sequencing revealed over 300 known miRNAs packaged into myotube EVs. Many were differentially expressed after EPS, either positively or negatively. Muscle-important miRNAs were present (miR-206 was 4.8-fold more prevalent than any other miRNA). EV treatments improved myoblast migration and differentiation without a frequency-specific influence. Gene Ontology analysis based on differentially expressed miRNAs between control and EPS-EVs indicates an effect of EPS frequency on muscle EV signaling.
Keywords: extracellular vesicles; C2C12 myotubes; electrical pulse stimulation; micro-RNA cargo extracellular vesicles; C2C12 myotubes; electrical pulse stimulation; micro-RNA cargo

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MDPI and ACS Style

Hingle, J.S.; McColl, R.S.; Vechetti, I.J.; Myburgh, K.H. Small Extracellular Vesicle Release Following Electrical Pulse Stimulation of C2C12 Myotubes: Effects on microRNA Cargo and Myoblast Migration and Differentiation. Int. J. Mol. Sci. 2026, 27, 4320. https://doi.org/10.3390/ijms27104320

AMA Style

Hingle JS, McColl RS, Vechetti IJ, Myburgh KH. Small Extracellular Vesicle Release Following Electrical Pulse Stimulation of C2C12 Myotubes: Effects on microRNA Cargo and Myoblast Migration and Differentiation. International Journal of Molecular Sciences. 2026; 27(10):4320. https://doi.org/10.3390/ijms27104320

Chicago/Turabian Style

Hingle, John S., Rhys S. McColl, Ivan J. Vechetti, and Kathryn H. Myburgh. 2026. "Small Extracellular Vesicle Release Following Electrical Pulse Stimulation of C2C12 Myotubes: Effects on microRNA Cargo and Myoblast Migration and Differentiation" International Journal of Molecular Sciences 27, no. 10: 4320. https://doi.org/10.3390/ijms27104320

APA Style

Hingle, J. S., McColl, R. S., Vechetti, I. J., & Myburgh, K. H. (2026). Small Extracellular Vesicle Release Following Electrical Pulse Stimulation of C2C12 Myotubes: Effects on microRNA Cargo and Myoblast Migration and Differentiation. International Journal of Molecular Sciences, 27(10), 4320. https://doi.org/10.3390/ijms27104320

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