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dc.contributor.authorHutcheson, Joshua D.en_US
dc.contributor.authorGoettsch, Claudiaen_US
dc.contributor.authorPham, Tanen_US
dc.contributor.authorIwashita, Masayaen_US
dc.contributor.authorAikawa, Masanorien_US
dc.contributor.authorSingh, Sasha A.en_US
dc.contributor.authorAikawa, Elenaen_US
dc.date.accessioned2015-01-05T18:27:54Z
dc.date.issued2014en_US
dc.identifier.citationHutcheson, Joshua D., Claudia Goettsch, Tan Pham, Masaya Iwashita, Masanori Aikawa, Sasha A. Singh, and Elena Aikawa. 2014. “Enrichment of calcifying extracellular vesicles using density-based ultracentrifugation protocol.” Journal of Extracellular Vesicles 3 (1): 10.3402/jev.v3.25129. doi:10.3402/jev.v3.25129. http://dx.doi.org/10.3402/jev.v3.25129.en
dc.identifier.issn2001-3078en
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:13581174
dc.description.abstractCalcifying extracellular vesicles (EVs) released from cells within atherosclerotic plaques have received increased attention for their role in mediating vascular calcification, a major predictor of cardiovascular morbidity and mortality. However, little is known about the difference between this pathologic vesicle population and other EVs that contribute to physiological cellular processes. One major challenge that hinders research into these differences is the inability to selectively isolate calcifying EVs from other vesicle populations. In this study, we hypothesized that the formation of mineral within calcifying EVs would increase the density of the vesicles such that they would pellet at a faster rate during ultracentrifugation. We show that after 10 min of ultracentrifugation at 100,000×g, calcifying EVs are depleted from the conditioned media of calcifying coronary artery smooth muscle cells and are enriched in the pelleted portion. We utilized mass spectrometry to establish functional proteomic differences between the calcifying EVs enriched in the 10 min ultracentrifugation compared to other vesicle populations preferentially pelleted by longer ultracentrifugation times. The procedures established in this study will allow us to enrich the vesicle population of interest and perform advanced proteomic analyses to find subtle differences between calcifying EVs and other vesicle populations that may be translated into therapeutic targets for vascular calcification. Finally, we will show that the differences in ultracentrifugation times required to pellet the vesicle populations can also be used to estimate physical differences between the vesicles.en
dc.language.isoen_USen
dc.publisherCo-Action Publishingen
dc.relation.isversionofdoi:10.3402/jev.v3.25129en
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC4261240/pdf/en
dash.licenseLAAen_US
dc.subjectcalcificationen
dc.subjectextracellular vesiclesen
dc.subjectatherosclerosisen
dc.subjectultracentrifugationen
dc.subjectisolationen
dc.titleEnrichment of calcifying extracellular vesicles using density-based ultracentrifugation protocolen
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden
dc.relation.journalJournal of Extracellular Vesiclesen
dash.depositing.authorHutcheson, Joshua D.en_US
dc.date.available2015-01-05T18:27:54Z
dc.identifier.doi10.3402/jev.v3.25129*
dash.authorsorderedfalse
dash.contributor.affiliatedIwashita, Masaya
dash.contributor.affiliatedGoettsch, Claudia
dash.contributor.affiliatedAikawa, Masanori
dash.contributor.affiliatedHutcheson, Joshua
dash.contributor.affiliatedSingh, Sasha
dash.contributor.affiliatedAikawa, Elena


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