Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes

Significance The last decade has seen a rapid expansion of interest in extracellular vesicles (EVs), proposed to mediate cell–cell communication in patho/physiological conditions. Although heterogeneity of EVs has become obvious, as highlighted recently by the International Society for Extracellular Vesicles, the field is lacking specific tools to distinguish EVs of different intracellular origins, and thus probably different functions. Here, thanks to a comprehensive comparison of different types of EVs isolated from a single cell type, we define proteins generically present in EVs, small EV-specific and -excluded ones, and a few specific of endosome-derived exosomes or nonexosomal small EVs. This work will allow proper evaluation of the molecular mechanisms of biogenesis and secretion and the respective functions of subtypes of EVs.