Techniques used for the isolation and characterization of extracellular vesicles: results of a worldwide survey

TLDR

Extracellular vesicles are key mediators of intercellular communication, yet the field has rapidly expanded with many poorly standardized isolation and characterization techniques, and their heterogeneous size, origin, and molecular composition make current practices largely unknown. The authors conducted a comprehensive survey of global EV isolation and characterization practices. The survey revealed that conditioned cell culture media were used by 83% of respondents, ultracentrifugation was the most common isolation method (81%) with 59% employing combinations, only 9% used a single characterization method, and sample volume, type, and downstream application influenced technique choice.

Abstract

Extracellular vesicles (EVs) represent an important mode of intercellular communication. Research in this field has grown rapidly in the last few years, and there is a plethora of techniques for the isolation and characterization of EVs, many of which are poorly standardized. EVs are heterogeneous in size, origin and molecular constituents, with considerable overlap in size and phenotype between different populations of EVs. Little is known about current practices for the isolation, purification and characterization of EVs. We report here the first large, detailed survey of current worldwide practices for the isolation and characterization of EVs. Conditioned cell culture media was the most widely used material (83%). Ultracentrifugation remains the most commonly used isolation method (81%) with 59% of respondents use a combination of methods. Only 9% of respondents used only 1 characterization method, with others using 2 or more methods. Sample volume, sample type and downstream application all influenced the isolation and characterization techniques employed.