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

By C. Gardiner, D. D. Vizio, S. Sahoo, C. Théry, Kenneth W. Witwer, M. Wauben, Andrew F. Hill. Journal of Extracellular Vesicles, 2016

938 citations recorded by the source corpus.

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.

Subfields: Cancer Research, Immunology and Allergy, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/6679d8ecd6407a553ede0ed6eb8690bcb80931ee

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Citations in the corpus

A Review of Exosomal Isolation Methods: Is Size Exclusion Chromatography the Best Option? (2020, background): “Nonetheless, a worldwide survey [56] from one hundred and ninety-six members of ISEV was collected from an online questionnaire administered via email in October 2015 [56].”
Obstacles and opportunities in the functional analysis of extracellular vesicle RNA – an ISEV position paper (2017, neutral): “Pros and cons of commonly used EV isolation techniques have been reviewed more elaborately elsewhere (see for example [29,30]), but are briefly summarised below.”
Methods for Separation and Characterization of Extracellular Vesicles: Results of a Worldwide Survey Performed by the ISEV Rigor and Standardization Subcommittee (2020, methodology): “[8], cell culture-conditioned serum-free medium is the most common source of EVs.”
Plant exosome like nanovesicles: Emerging therapeutics and drug delivery nanoplatforms. (2020, methodology): “However, this technique has been reported to co-precipitate other EVs alongside ELNVs as well as RNA-protein complexes.(40) Re-engineering of PELNVs”
Advances in therapeutic applications of extracellular vesicles (2019, methodology): “IMPACT OF ISOLATION METHODS ON EV INTEGRITY AND PURITY The current standard technique for EV isolation is differential centrifugation with ultracentrifugation, encompassing a series of centrifugations to remove floating cells and cellular debris, followed by ultracentrifugation to pellet EVs (11, 68”
Exosomes as a new frontier of cancer liquid biopsy (2022, methodology): “Exosome characterization As suggested in the Minimal information for studies of extracellular vesicles 2018 (MISEV2018), the identification of exosomes should include western blot verification of exosome-specific markers and at least two methods for characterization of single exosome [94, 95].”
Manufacturing Exosomes: A Promising Therapeutic Platform. (2018, neutral): “This was evident in a worldwide survey that showed that 77% of respondents used less than 100 ml of starting material, despite 83% of researchers using material generated from cell cultures [43].”