| Author | Arteaga-Blanco, Luis A. | |
| Author | Mojoli, Andrés | |
| Author | Monteiro, Robson Q. | |
| Author | Sandim, Vanessa | |
| Author | Menna-Barreto, Rubem F. S. | |
| Author | Dutra, Filipe Santos Dutra | |
| Author | Bozza, Patrícia T. | |
| Author | Resende, Rafael de Oliveira | |
| Author | Bou-Habib, Dumith Chequer | |
| Access date | 2020-08-31T20:44:35Z | |
| Available date | 2020-08-31T20:44:35Z | |
| Document date | 2020 | |
| Citation | ARTEAGA-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. | pt_BR |
| ISSN | 1932-6203 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/43093 | |
| Language | eng | pt_BR |
| Publisher | Public Library of Science | pt_BR |
| Rights | open access | |
| Subject in Portuguese | Vesículas extracelulares | pt_BR |
| Subject in Portuguese | Macrófagos primários humanos | pt_BR |
| Subject in Portuguese | Monócitos circulantes | pt_BR |
| Subject in Portuguese | Internalização | pt_BR |
| Title | Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes | pt_BR |
| Type | Article | |
| DOI | 10.1371/journal.pone.0237795 | |
| Abstract | Extracellular 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. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa sobre o Timo. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa sobre o Timo. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Celular. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunofarmacologia. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunofarmacologia. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa sibre o Timo. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa sibre o Timo. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciências e Tecnologia em Neuroimunomodulação. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Subject | Internalization | pt_BR |
| Subject | Small extracellular vesicles | pt_BR |
| Subject | Human primary macrophages | pt_BR |
| Subject | Circulating monocytes | pt_BR |
