Techniques used for the isolation and characterization of extracellular vesicles: results of a worldwide survey.
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.
Haemostasis Research Unit, Research Department of Haematology, University College London, London, UK; c.gardiner@ucl.ac.uk.
Chris
C
Gardiner
Icahn School of Medicine, Cardiovascular Research Center, Cedars-Sinai, Los Angeles, CA, USA.
Dolores
D
Di Vizio
Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
Susmita
S
Sahoo
Institut Curie, PSL Research University, INSERM U932, Paris, France.
Clotilde
C
Théry
Johns Hopkins University School of Medicine, Baltimore, USA.
Kenneth W
KW
Witwer
Department of Biochemistry and Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
Marca
M
Wauben
Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Australia.
Andrew F
AF
Hill
RNA analysis; characterization; extracellular vesicles; flow cytometry; isolation; proteomics; purification; single vesicle analysis