High-speed centrifugation induces aggregation of extracellular vesicles

Abstract

Plasma and other body fluids contain cell-derived extracellular vesicles (EVs), which participate in physiopathological processes and have potential biomedical applications. In order to isolate, concentrate and purify EVs, high-speed centrifugation is often used. We show here, using electron microscopy, receptor-specific gold labelling and flow cytometry, that high-speed centrifugation induces the formation of EV aggregates composed of a mixture of EVs of various phenotypes and morphologies. The presence of aggregates made of EVs of different phenotypes may lead to erroneous interpretation concerning the existence of EVs harbouring surface antigens from different cell origins.

• extracellular vesicles
• blood plasma
• cryo-electron microscopy
• immuno-gold electron microscopy
• flow cytometry
• vesicle aggregation
• centrifugation

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Acknowledgements

We thank Mrs. Chaléat (Laboratoire Mutualiste d'Analyses Médicales, Pessac, France) for her help with the collection of blood samples. We thank Dr. Chris Gardiner (University College London, UK) for helpful discussions.

Notes

To access the supplementary material to this article, please see Supplementary files under ‘Article Tools’.