Extracellular vesicles (EVs) are membranous particles released from all organisms and cell types. EVs carry proteins, RNAs, microRNAs, and metabolites that, when released, can be taken up by distant cells or tissues, providing a means for cell-to-cell communication and organ cross-talk. Apoptotic bodies, exosomes, microvesicles, and microparticles are subtypes of extracellular vesicles. Isolated EVs are classified as a distinct subtype (e.g., exosomes) if the source cellular compartments or organelles can be confirmed. Otherwise, EVs can be classified by their physical characteristics: size, density, presence of hallmark EV proteins, and cellular origins.

In light of EVs' increasing interest and reporting in a variety of research fields, it is necessary to ensure the fidelity of methods used to isolate EVs. EV isolation and enrichment are challenging for various reasons, including the high likelihood of contamination from culture media constituents, cellular debris, and lipoproteins if isolated from plasma. EVs isolated from other biofluids, including urine, cerebrospinal fluid, and bronchoalveolar lavage, also present unique challenges.

The goal of this Methods Collection is to provide protocols for EV isolation that adhere to the minimum standards for EV research (MISEV 2018) and that profile EV constituents using the omics platform. Methods optimized for one or more critical steps in EV isolation and reporting, such as collection, storage, isolation, and characterization, are welcome for submissions. This collection will provide researchers with a toolbox that can be sorted as needed to acquire optimal and high-quality EVs.