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. 2016 Oct 31:5:32945.
doi: 10.3402/jev.v5.32945. eCollection 2016.

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

Chris Gardiner  1 , Dolores Di Vizio  2 , Susmita Sahoo  3 , Clotilde Théry  4 , Kenneth W Witwer  5 , Marca Wauben  6 , Andrew F Hill  7
Affiliations

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

Chris Gardiner et al. J Extracell Vesicles. .

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.

Keywords: RNA analysis; characterization; extracellular vesicles; flow cytometry; isolation; proteomics; purification; single vesicle analysis.

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Conflict of interest statement

and funding The authors have not received any funding or benefits from industry or elsewhere to conduct this study.

Figures

Fig. 1
Fig. 1
Respondents by country (%).
Fig. 2
Fig. 2
(a) Starting material used by respondents by percentage. (b) Starting volume by sample type.Only responses indicating a single type of sample were included in the analysis (0–5 ml, n = 22; 5–100 ml, n = 51; >100 ml, n = 28).
Fig. 3
Fig. 3
(a) Primary isolation method (% of respondents) and (b) isolation method by starting volume of material.
Fig. 4
Fig. 4
Methods used for EV characterization (%).
Fig. 5
Fig. 5
Downstream applications (%).
See this image and copyright information in PMC

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