Isolation and characterization of extracellular vesicles produced by cell lines

Cells produce two broad classes of extracellular vesicles (EVs), exosomes and microvesicles (MVs). Exosomes are 30–150 nm vesicles derived from multivesicular bodies, while MVs are 200–1,000 nm vesicles that pinch off from plasma membranes. Reliable isolation of EVs is crucial to understand their biochemical and functional properties. Here, we describe a protocol to isolate and characterize EVs from conditioned medium from mammalian cell lines. This protocol has been optimized for adherent cells but can also be adapted for suspension cells. For complete details on the use and execution of this protocol, please refer to Latifkar et al. (2019).