Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2020 |
| Outros Autores: | , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/43093 |
Resumo: | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa sobre o Timo. Rio de Janeiro, RJ, Brasil. |
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Arteaga-Blanco, Luis A.Mojoli, AndrésMonteiro, Robson Q.Sandim, VanessaMenna-Barreto, Rubem F. S.Dutra, Filipe Santos DutraBozza, Patrícia T.Resende, Rafael de OliveiraBou-Habib, Dumith Chequer2020-08-31T20:44:35Z2020-08-31T20:44:35Z2020ARTEAGA-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. 2020.1932-6203https://www.arca.fiocruz.br/handle/icict/4309310.1371/journal.pone.0237795engPublic Library of ScienceVesículas extracelularesMacrófagos primários humanosMonócitos circulantesInternalizaçãoInternalizationSmall extracellular vesiclesHuman primary macrophagesCirculating monocytesCharacterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa sobre o Timo. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa sobre o Timo. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Celular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunofarmacologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunofarmacologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa sibre o Timo. Rio de Janeiro, RJ, Brasil.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.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.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; 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| dc.title.pt_BR.fl_str_mv |
Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes |
| title |
Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes |
| spellingShingle |
Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes Arteaga-Blanco, Luis A. Vesículas extracelulares Macrófagos primários humanos Monócitos circulantes Internalização Internalization Small extracellular vesicles Human primary macrophages Circulating monocytes |
| title_short |
Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes |
| title_full |
Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes |
| title_fullStr |
Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes |
| title_full_unstemmed |
Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes |
| title_sort |
Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes |
| author |
Arteaga-Blanco, Luis A. |
| author_facet |
Arteaga-Blanco, Luis A. Mojoli, Andrés Monteiro, Robson Q. Sandim, Vanessa Menna-Barreto, Rubem F. S. Dutra, Filipe Santos Dutra Bozza, Patrícia T. Resende, Rafael de Oliveira Bou-Habib, Dumith Chequer |
| author_role |
author |
| author2 |
Mojoli, Andrés Monteiro, Robson Q. Sandim, Vanessa Menna-Barreto, Rubem F. S. Dutra, Filipe Santos Dutra Bozza, Patrícia T. Resende, Rafael de Oliveira Bou-Habib, Dumith Chequer |
| author2_role |
author author author author author author author author |
| dc.contributor.author.fl_str_mv |
Arteaga-Blanco, Luis A. Mojoli, Andrés Monteiro, Robson Q. Sandim, Vanessa Menna-Barreto, Rubem F. S. Dutra, Filipe Santos Dutra Bozza, Patrícia T. Resende, Rafael de Oliveira Bou-Habib, Dumith Chequer |
| dc.subject.other.pt_BR.fl_str_mv |
Vesículas extracelulares Macrófagos primários humanos Monócitos circulantes Internalização |
| topic |
Vesículas extracelulares Macrófagos primários humanos Monócitos circulantes Internalização Internalization Small extracellular vesicles Human primary macrophages Circulating monocytes |
| dc.subject.en.pt_BR.fl_str_mv |
Internalization Small extracellular vesicles Human primary macrophages Circulating monocytes |
| description |
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa sobre o Timo. Rio de Janeiro, RJ, Brasil. |
| publishDate |
2020 |
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2020-08-31T20:44:35Z |
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2020-08-31T20:44:35Z |
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2020 |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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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. |
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https://www.arca.fiocruz.br/handle/icict/43093 |
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1932-6203 |
| dc.identifier.doi.none.fl_str_mv |
10.1371/journal.pone.0237795 |
| identifier_str_mv |
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. 1932-6203 10.1371/journal.pone.0237795 |
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