Comprehensive evaluation of methods for small extracellular vesicles separation from human plasma, urine and cell culture medium

One of the challenges that restricts the evolving extracellular vesicle (EV) research field is the lack of a consensus method for EV separation. This may also explain the diversity of the experimental results, as co‐separated soluble proteins and lipoproteins may impede the interpretation of experim...

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Vydané v:Journal of extracellular vesicles Ročník 10; číslo 2; s. e12044 - n/a
Hlavní autori: Dong, Liang, Zieren, Richard C., Horie, Kengo, Kim, Chi‐Ju, Mallick, Emily, Jing, Yuezhou, Feng, Mingxiao, Kuczler, Morgan D., Green, Jordan, Amend, Sarah R., Witwer, Kenneth W., de Reijke, Theo M., Cho, Yoon‐Kyoung, Pienta, Kenneth J., Xue, Wei
Médium: Journal Article
Jazyk:English
Vydavateľské údaje: United States John Wiley & Sons, Inc 01.12.2020
John Wiley and Sons Inc
Predmet:
Biofiltration
Cell culture
Chromatography, Gel - methods
Contaminants
Culture media
Culture Media - chemistry
EV yield
Extracellular vesicles
Extracellular Vesicles - metabolism
Humans
Invoices
Lipoproteins
Male
methods comparison
Microfluidics
Phenotyping
Plasma
Plasma - chemistry
Plasma - metabolism
Prostatic Neoplasms - metabolism
Reagents
sample purity
single particle phenotyping
small extracellular vesicle separation
Tumor Cells, Cultured
Ultracentrifugation
Ultracentrifugation - methods
Ultrafiltration
Urine
Urine - chemistry
United States > US
EV yield
sample purity
methods comparison
small extracellular vesicle separation
single particle phenotyping
ISSN:2001-3078, 2001-3078
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Shrnutí:One of the challenges that restricts the evolving extracellular vesicle (EV) research field is the lack of a consensus method for EV separation. This may also explain the diversity of the experimental results, as co‐separated soluble proteins and lipoproteins may impede the interpretation of experimental findings. In this study, we comprehensively evaluated the EV yields and sample purities of three most popular EV separation methods, ultracentrifugation, precipitation and size exclusion chromatography combined with ultrafiltration, along with a microfluidic tangential flow filtration device, Exodisc, in three commonly used biological samples, cell culture medium, human urine and plasma. Single EV phenotyping and density‐gradient ultracentrifugation were used to understand the proportion of true EVs in particle separations. Our findings suggest Exodisc has the best EV yield though it may co‐separate contaminants when the non‐EV particle levels are high in input materials. We found no 100% pure EV preparations due to the overlap of their size and density with many non‐EV particles in biofluids. Precipitation has the lowest sample purity, regardless of sample type. The purities of the other techniques may vary in different sample types and are largely dependent on their working principles and the intrinsic composition of the input sample. Researchers should choose the proper separation method according to the sample type, downstream analysis and their working scenarios.
Bibliografia:ObjectType-Article-1
SourceType-Scholarly Journals-1
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Kenneth J. Pienta, Wei Xue share the co‐senior authorship.
Liang Dong, Richard C. Zieren, and Kengo Horie contributed equally to this work.
ISSN:2001-3078
2001-3078
DOI:10.1002/jev2.12044