Extracellular Vesicle Isolation and Purification from Skeletal Muscle Cells.

Extracellular vesicles (EVs) have emerged as critical mediators of intercellular communication and hold considerable promise as biomarkers and therapeutic vectors. In skeletal muscle, EVs transport proteins, RNA, and other bioactive molecules, such as myokines, making them valuable indicators of muscle physiology and pathology. This method paper presents a detailed protocol for the isolation, purification, and characterization of EVs derived from human myoblasts. Our optimized workflow ensures high yield and sample quality, enabling comprehensive downstream molecular analysis. Additionally, we describe advanced techniques for assessing EV physical characteristics, concentration, and cargo content, providing an essential toolkit for investigating skeletal muscle EV biology and potential role in disease diagnostics. This protocol facilitates the efficient exploration of skeletal muscle-derived EVs and their implications in muscle function and pathology.