Protocol for endothelial cell-derived extracellular vesicle isolation and characterization.

Endothelial cells regulate intercellular and inter-organ communication through extracellular vesicles (EVs), which have diagnostic and therapeutic potential. We describe single-particle interferometric reflectance imaging directly from culture supernatants, followed by ultracentrifugation isolation of EVs. We determine EV size and concentration by nanoparticle tracking analysis and characterize EVs by western blotting and transmission electron microscopy. This protocol can be applied to other cell systems of EV biology. For details on the execution of this protocol, please refer to Timms et al.1.