Mode of induction of platelet-derived extracellular vesicles is a critical determinant of their phenotype and function

Ferreira, P., Bozbas, E., Tannetta, D., Alroqaiba, N., Zhou, R., Crawley, J., Gibbins, J. ORCID: https://orcid.org/0000-0002-0372-5352, Jones, C. ORCID: https://orcid.org/0000-0001-7537-1509, Ahnstrom, J. and Yaqoob, P. ORCID: https://orcid.org/0000-0002-6716-7599 (2020) Mode of induction of platelet-derived extracellular vesicles is a critical determinant of their phenotype and function. Scientific Reports, 10 (1). 18061. ISSN 2045-2322 doi: 10.1038/s41598-020-73005-3

Abstract/Summary

Platelet-derived extracellular vesicles (PDEVs) are the most abundant amongst all types of EVs in the circulation. However, the mechanisms leading to PDEVs release, their role in coagulation and phenotypic composition are poorly understood. PDEVs from washed platelets were generated using different stimuli and were characterised using nanoparticle tracking analysis. Procoagulant properties were evaluated by fluorescence flow cytometry and calibrated automated thrombography. EVs from plasma were isolated and concentrated using a novel protocol involving a combination of size exclusion chromatography and differential centrifugation, which produces pure and concentrated EVs. Agonist stimulation enhanced PDEV release, but did not alter the average size of EVs compared to those produced by unstimulated platelets. Agonist stimulation led to lower negatively-charged phospholipid externalization in PDEVs, which was reflected in the lower procoagulant activity compared to those generated without agonist stimulation. Circulating EVs did not have externalized negatively-charged phospholipids. None of the 4 types of EVs presented tissue factor. The mechanism by which PDEV formation is induced is a critical determinant of its phenotype and function. Importantly, we have developed methods to obtain clean, concentrated and functional EVs derived from platelet-free plasma and washed platelets, which can be used to provide novel insight into their biological functions.

Item Type	Article
URI	https://reading-clone.eprints-hosting.org/id/eprint/92496
Identification Number/DOI	10.1038/s41598-020-73005-3
Refereed	Yes
Divisions	Interdisciplinary centres and themes > Institute for Cardiovascular and Metabolic Research (ICMR) Life Sciences > School of Chemistry, Food and Pharmacy > Department of Food and Nutritional Sciences > Human Nutrition Research Group
Publisher	Nature Publishing Group
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