期刊
JOURNAL OF EXTRACELLULAR VESICLES
卷 9, 期 1, 页码 -出版社
TAYLOR & FRANCIS LTD
DOI: 10.1080/20013078.2020.1791450
关键词
Extracellular vesicles; exosomes; isolation; lipoprotein particles; human plasma
类别
资金
- China Scholarship Council [201606300042]
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek [184.032.201]
- ERC Consolidator Grant [724425]
- European Research Council (ERC) [724425] Funding Source: European Research Council (ERC)
Extracellular vesicles (EV) are membrane encapsulated nanoparticles that can function in intercellular communication, and their presence in biofluids can be indicative for (patho)physiological conditions. Studies aiming to resolve functionalities of EV or to discover EV-associated biomarkers for disease in liquid biopsies are hampered by limitations of current protocols to isolate EV from biofluids or cell culture medium. EV isolation is complicated by the >10(5)-fold numerical excess of other types of particles, including lipoproteins and protein complexes. In addition to persisting contaminants, currently available EV isolation methods may suffer from inefficient EV recovery, bias for EV subtypes, interference with the integrity of EV membranes, and loss of EV functionality. In this study, we established a novel three-step non-selective method to isolate EV from blood or cell culture media with both high yield and purity, resulting in 71% recovery and near to complete elimination of unrelated (lipo)proteins. This EV isolation procedure is independent of ill-defined commercial kits, and apart from an ultracentrifuge, does not require specialised expensive equipment.
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