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Extracellular vesicles from blood plasma: determination of their morphology, size, phenotype and concentration.

hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorARRAUD, Nicolas
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorLINARES, Romain
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorTAN, Sisareuth
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorGOUNOU, Céline
hal.structure.identifierHématopoïèse Leucémique et Cibles Thérapeutiques
dc.contributor.authorPASQUET, Jean-Max
hal.structure.identifierInstitut de Chimie de la Matière Condensée de Bordeaux [ICMCB]
dc.contributor.authorMORNET, Stéphane
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorBRISSON, Alain R.
dc.date.issued2014
dc.identifier.issn1538-7933
dc.description.abstractEnBACKGROUND: Plasma and other body fluids contain membranous extracellular vesicles (EVs), which are considered to derive from activated or apoptotic cells. EVs participate in physiological and pathological processes and have potential applications in diagnostics or therapeutics. Knowledge on EVs is, however, limited, mainly due to their sub-micrometer size and to intrinsic limitations in methods applied for their characterization. OBJECTIVES: Our aim was to provide a comprehensive description of EVs from plasma of healthy subjects. METHODS: Cryo-transmission electron microscopy combined with receptor-specific gold labeling was used to reveal the morphology, size and phenotype of EVs. An original approach based on sedimentation on electron microscopy grids was developed for enumerating EVs. A correlation was performed between conventional flow cytometry and electron microscopy results. RESULTS: We show that platelet-free plasma samples contain spherical EVs, 30 nm to 1 μm in diameter, tubular EVs, 1-5 μm long, and membrane fragments, 1-8 μm large. We show that only a minority of EVs expose the procoagulant lipid phosphatidylserine, in contrast to the classical theory of EV formation. In addition, the concentrations of the main EV sub-populations are determined after sedimentation on EM grids. Finally, we show that conventional flow cytometry, the main method of EV characterization, detects only about 1% of them. CONCLUSION: This study brings novel insights on EVs from normal plasma and provides a reference for further studies of EVs in disease situations.
dc.language.isoen
dc.publisherWiley
dc.subject.enBlood plasma
dc.subject.enCell-derived microparticles
dc.subject.enCryo-electron microscopy
dc.subject.enFlow cytometry
dc.subject.enImmunogold techniques
dc.title.enExtracellular vesicles from blood plasma: determination of their morphology, size, phenotype and concentration.
dc.typeArticle de revue
dc.identifier.doi10.1111/jth.12554
dc.subject.halSciences du Vivant [q-bio]/Médecine humaine et pathologie/Hématologie
dc.subject.halSciences du Vivant [q-bio]/Ingénierie biomédicale/Imagerie
bordeaux.journalJournal of Thrombosis and Haemostasis
bordeaux.page614-627
bordeaux.volume12
bordeaux.issue5
bordeaux.peerReviewedoui
hal.identifierhal-00996592
hal.version1
hal.popularnon
hal.audienceInternationale
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-00996592v1
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