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Protocol to isolate endosomal and small extracellular vesicles from cultured cells through ultracentrifugation

2026-06-19Artigo científico Acesso aberto
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dc.contributor.authorDomingues, Maria Nolasco
dc.contributor.authorPalhinhas, Luís
dc.contributor.authorPereira, Paulo
dc.contributor.authorFerreira, João Vasco
dc.contributor.institutionNOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)
dc.contributor.institutioniNOVA4Health - pólo NMS
dc.contributor.pblCell Press
dc.date.accessioned2026-06-26T16:23:02Z
dc.date.available2026-06-26T16:23:02Z
dc.date.issued2026-06-19
dc.descriptionPublisher Copyright: © 2026 The Authors.
dc.description.abstractHere, we present a protocol to isolate endosomal fractions using sucrose-density gradient ultracentrifugation and to recover small extracellular vesicles (sEVs), enriched in exosomes, using sequential ultracentrifugation from mammalian cell lines. This combined approach enables the separation and analysis of early endosome (EE), late endosome (LE), and sEV fractions. We provide detailed procedures for cell culture preparation, conditioned media collection, differential centrifugation, gradient layering, and fraction purification, facilitating downstream characterization and functional assays. For complete details on the use and execution of this protocol, please refer to Ferreira et al.en
dc.description.versionpublishersversion
dc.description.versionpublished
dc.format.extent2669980
dc.identifier.doi10.1016/j.xpro.2026.104531
dc.identifier.issn2666-1667
dc.identifier.otherPURE: 163159536
dc.identifier.otherPURE UUID: 9553fffc-ce64-4f7d-a6ed-dae9359b46b8
dc.identifier.otherScopus: 105037400461
dc.identifier.otherORCID: /0000-0002-9908-2290/work/218920305
dc.identifier.urihttp://hdl.handle.net/10362/204134
dc.identifier.urlhttps://www.scopus.com/pages/publications/105037400461
dc.language.isoeng
dc.peerreviewedyes
dc.subjectGeneral Neuroscience
dc.subjectGeneral Immunology and Microbiology
dc.subjectGeneral Biochemistry,Genetics and Molecular Biology
dc.titleProtocol to isolate endosomal and small extracellular vesicles from cultured cells through ultracentrifugationen
dc.typejournal article
degois.publication.issue2
degois.publication.titleSTAR Protocols
degois.publication.volume7
dspace.entity.typePublication
rcaap.rightsopenAccess

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