We discovered that a protein called Wnt7a is elevated and released from newly regenerating muscle fibers in response to muscle injury, triggering a healing response.

However, isolating extracellular vesicles (EVs) from muscle tissue is challenging. Traditional methods, like ultracentrifugation, often lead to contamination with non-EV proteins and cellular fragments.

To overcome this, we developed a new protocol using Tangential Flow Filtration (TFF) and Size Exclusion Chromatography (SEC). This method allows us to purify Wnt7a associated with EVs (retentate fraction) and free non-EV Wnt7a (permeate fraction).

Our optimized protocol removes large EVs through centrifugation, concentrates and washes EVs with TFF, and eliminates remaining soluble proteins with SEC.

This approach is effective for isolating EVs from muscle tissue without contamination, improving the reproducibility, efficiency, and purity of EV preparations. The purified EVs maintain structural integrity, making them suitable for various bioactivity and analytic assays in both in vitro and in vivo settings.