Isolation of Extracellular Vesicles by Ultracentrifugation

Extracellular vesicles (EVs) represent a group of heterogeneous vesicles that can be obtained from almost all biofluids. EVs, including microvesicles, exosomes, and apoptotic bodies , can deliver bioactive cargos and signaling molecules. Various physiological roles and pathophysiological roles for EVs in diseases such as cancer, infectious diseases, endocrine diseases, and neurodegenerative disorders have been recognized. These observations highlight EVs as potential novel biomarkers and targets for therapeutic intervention. One of the major limitations in the use of EVs for diagnosis and therapeutic purposes is the lack of standardization of isolation techniques. Here, we describe protocols for ultracentrifugation and sucrose gradient isolation methods, which are the current gold standard, and are the most studied methods for EV isolation.