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Comparison of serum exosome isolation methods on co-precipitated free microRNAs.
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2020 (English)In: PeerJ, E-ISSN 2167-8359, Vol. 8, article id e9434Article in journal (Refereed) Published
Abstract [en]

Background: Exosomes are nano-sized extracellular vesicles containing different biomolecules such as proteins and microRNAs (miRNAs) that mediate intercellular communication. Recently, numerous studies have reported the important functions of exosomal miRNAs in disease development and the potential clinical application as diagnostic biomarkers. Up to now, the most commonly used methods to extract exosomes are ultracentrifugation (UC) and precipitation-based commercial kit (e.g., ExoQuick). Generally, both UC and ExoQuick method could co-isolate contaminating proteins along with exosomes, with the UC method yielding even purer exosomes than ExoQuick. However, the comparison of these two methods on co-precipitated free miRNAs is still unknown.

Methods: In this study, we isolated exosomes from the human serum with exogenously added cel-miR-39 by UC and ExoQuick and compared the proportion of cel-miR-39 co-precipitated with exosomes extracted by these two methods.

Results: Using exogenous cel-miR-39 as free miRNAs in serum, we concluded that ExoQuick co-isolates a small proportion of free miRNAs while UC hardly precipitates any free miRNAs. We also found that incubation at 37 °C for 1 h could decrease the proportion of free miRNAs, and exosomal miRNAs like miR-126 and miR-152 also decreased when RNase A was used. In conclusion, our findings provide essential information about the details of serum exosome isolation methods for further research on exosomal miRNAs.

Place, publisher, year, edition, pages
PeerJ , 2020. Vol. 8, article id e9434
Keywords [en]
ExtraPEG, Isolation method, MicroRNAs, Serum, Ultracentrifugation, Exosome
National Category
Medical Laboratory and Measurements Technologies
Identifiers
URN: urn:nbn:se:kth:diva-283781DOI: 10.7717/peerj.9434ISI: 000563710000002PubMedID: 32923177Scopus ID: 2-s2.0-85091530390OAI: oai:DiVA.org:kth-283781DiVA, id: diva2:1475398
Note

QC 20201019

Available from: 2020-10-12 Created: 2020-10-12 Last updated: 2023-08-28Bibliographically approved

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Williams, Cecilia

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CiteExportLink to record
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Citation style
  • apa
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  • vancouver
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Language
  • de-DE
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  • fi-FI
  • nn-NO
  • nn-NB
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  • Other locale
More languages
Output format
  • html
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  • asciidoc
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