Related Experiment Video

Updated: Jun 15, 2025

Enzymatic Isolation of Skeletal Muscle Interstitial Extracellular Vesicles
08:50

Enzymatic Isolation of Skeletal Muscle Interstitial Extracellular Vesicles

Published on: February 7, 2025

849

Extracellular vesicles may provide an alternative detoxification pathway during skeletal muscle myoblast ageing

María Fernández-Rhodes1, Emma Buchan2, Stephanie D Gagnon1

  • 1School of Sport Exercise and Health Sciences, Loughborough University Loughborough UK.

Journal of Extracellular Biology
|August 22, 2024
PubMed

Insights

Human skeletal muscle extracellular vesicles (SM-EVs) change with age. Older adults

Area of Science:

  • Somatic Cell Therapeutics
  • Extracellular Vesicle Biology
  • Aging Research

Background:

  • Skeletal muscle (SM) functions as a secretory organ, releasing myokines and extracellular vesicles (SM-EVs) that influence myogenesis and homeostasis.
  • Previous studies have documented age-related changes in murine SM-EVs, but a comprehensive profile in human models is lacking.
  • This study aims to bridge this gap by comparing SM-EVs from young and old human primary skeletal muscle cells (HPMCs).

Purpose of the Study:

  • To provide the first comprehensive comparison of SM-EVs from young and old human primary skeletal muscle cells (HPMCs).
  • To map the changes in SM-EVs associated with skeletal muscle aging in humans.
  • To investigate the potential role of SM-EVs in homeostasis and detoxification during SM aging.

Main Methods:

  • Human primary skeletal muscle cells (HPMCs) were isolated from young (24 ± 1.7 years) and older (69 ± 2.6 years) participants.
  • Cells were immunomagnetically sorted for the myogenic marker CD56 (N-CAM) and cultured as pure or mixed populations.
  • SM-EVs were isolated using an optimized ultrafiltration and size exclusion chromatography (UF + SEC) protocol.
  • EVs were characterized using Raman spectroscopy (RS) and liquid chromatography-mass spectrometry (LC-MS).

Main Results:

  • Minimal variations in basic EV parameters (particle number, size, protein markers) were observed between young and old SM-EVs.
  • Raman spectroscopy revealed increased protein (amide I), lipid (phospholipids, phosphatidylcholine), and hypoxanthine signatures in older SM-EVs.
  • LC-MS identified 84 shared proteins, primarily involved in cell homeostasis, muscle maintenance, and transcriptional regulation.
  • Older SM-EVs were enriched in proteins associated with oxidative stress and DNA/RNA mutagenesis (e.g., TTC3, ICE1, ACACA).

Conclusions:

  • Skeletal muscle extracellular vesicles (SM-EVs) exhibit distinct biochemical profiles with aging in humans.
  • Aging SM-EVs show increased signatures of lipids, proteins, and hypoxanthine, alongside enrichment of oxidative stress and mutagenesis-related proteins.
  • These findings suggest that SM-EVs may play a role in cellular homeostasis and detoxification processes during skeletal muscle aging.
Keywords:
LC‐MS/MSRaman spectroscopyageingextracellular vesicleshuman primary cellsskeletal muscle

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