Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes

dc.creatorArteaga-Blanco, Luis A.
dc.creatorMojoli, Andrés
dc.creatorMonteiro, Robson Q.
dc.creatorSandim, Vanessa
dc.creatorMenna-Barreto, Rubem F. S.
dc.creatorDutra, Filipe Santos Dutra
dc.creatorBozza, Patrícia T.
dc.creatorResende, Rafael de Oliveira
dc.creatorBou-Habib, Dumith Chequer
dc.date2020-08-31T20:44:35Z
dc.date2020-08-31T20:44:35Z
dc.date2020
dc.date.accessioned2023-09-26T21:00:25Z
dc.date.available2023-09-26T21:00:25Z
dc.identifierARTEAGA-BLANCO, Luís A. et al. Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes. Plos One, v. 15, n. 8, e0237795, Aug. 2020.
dc.identifier1932-6203
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/43093
dc.identifier10.1371/journal.pone.0237795
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8867610
dc.descriptionExtracellular vesicles (EVs) are small membrane-limited structures derived from outward budding of the plasma membrane or endosomal system that participate in cellular communication processes through the transport of bioactive molecules to recipient cells. To date, there are no published methodological works showing step-by-step the isolation, characterization and internalization of small EVs secreted by human primary macrophages derived from circulating monocytes (MDM-derived sEVs). Thus, here we aimed to provide an alternative protocol based on differential ultracentrifugation (dUC) to describe small EVs (sEVs) from these cells. Monocyte-derived macrophages were cultured in EV-free medium during 24, 48 or 72 h and, then, EVs were isolated from culture supernatants by (dUC). Macrophages secreted a large amount of sEVs in the first 24 h, with size ranging from 40-150 nm, peaking at 105 nm, as evaluated by nanoparticle tracking analysis and scanning electron microscopy. The markers Alix, CD63 and CD81 were detected by immunoblotting in EV samples, and the co-localization of CD63 and CD81 after sucrose density gradient ultracentrifugation (S-DGUC) indicated the presence of sEVs from late endosomal origin. Confocal fluorescence revealed that the sEVs were internalized by primary macrophages after three hours of co-culture. The methodology here applied aims to contribute for enhancing reproducibility between the limited number of available protocols for the isolation and characterization of MDM-derived sEVs, thus providing basic knowledge in the area of EV methods that can be useful for those investigators working with sEVs released by human primary macrophages derived from circulating monocytes.
dc.formatapplication/pdf
dc.languageeng
dc.publisherPublic Library of Science
dc.rightsopen access
dc.subjectVesículas extracelulares
dc.subjectMacrófagos primários humanos
dc.subjectMonócitos circulantes
dc.subjectInternalização
dc.subjectInternalization
dc.subjectSmall extracellular vesicles
dc.subjectHuman primary macrophages
dc.subjectCirculating monocytes
dc.titleCharacterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes
dc.typeArticle


Este ítem pertenece a la siguiente institución