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Morphological and electrical disturbances after split-flow fractionation in murine macrophages

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dc.creatorUrbina Bonilla, Adriana del Pilarspa
dc.creatorGodoy-Silva R.spa
dc.creatorHoyos M.spa
dc.creatorCamacho M.spa
dc.date.accessioned2020-05-25T23:55:59Z
dc.date.available2020-05-25T23:55:59Z
dc.date.created2019spa
dc.description.abstractSplit-flow fractionation (SPLITT) is a family of techniques that separates in the absence of labeling using very low flow rates and force fields, and is therefore expected to minimize cell damage. Although it has been documented that separation methods cause physiological changes in immune cells that are attributable to mechanical stress and antibody labeling, SPLITT has not yet been examined for possible damaging effects of hydrodynamic stress, partly because it is assumed that the low flow rates and weak forces used in this technique do not generate significant mechanical stress. The aim of this study was to investigate the effects of SPLITT on cell function of a murine macrophage cell, and to compare these effects with those induced by centrifugation. Macrophages J774.2 were cultured in RPMI-enriched media, then detached from the culture flask and resuspended for 12 h. Cell suspensions were diluted in a buffered saline solution and exposed to SPLITT (flow rates 1–10 ml/min) or centrifugation (100–1500g) for 10 min. Cell viability, diameter, membrane potential, and nitric oxide production were measured. Under the operating conditions employed, cell viability was above 98% after SPLITT and centrifugation but cells suffered immediate hydrodynamic cell damage, including decreased cell diameter and membrane hyperpolarization which was inhibitable by 4-aminopyridine; nitric oxide production was not affected. Pressure values during SPLITT and centrifugation correlated with diameter and membrane potential. Our data do not support the assumption that SPLITT is innocuous to cell function. Some changes in SPLITT channel design are suggested to minimize cell damage. Membrane potential and cell diameter are sensitive indicators for the evaluation of sublethal damage in different cell models, and allow identification of optimal operating conditions on different scales. © 2019 Elsevier B.V.eng
dc.format.mimetypeapplication/pdf
dc.identifier.doihttps://doi.org/10.1016/j.chroma.2019.01.005
dc.identifier.issn219673
dc.identifier.urihttps://repository.urosario.edu.co/handle/10336/22287
dc.language.isoengspa
dc.publisherElsevier B.V.spa
dc.relation.citationEndPage112
dc.relation.citationStartPage104
dc.relation.citationTitleJournal of Chromatography A
dc.relation.citationVolumeVol. 1590
dc.relation.ispartofJournal of Chromatography A, ISSN:219673, Vol.1590,(2019); pp. 104-112spa
dc.relation.urihttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85059515678&doi=10.1016%2fj.chroma.2019.01.005&partnerID=40&md5=6567e52b5af03e42e05a679c18d4806bspa
dc.rights.accesRightsinfo:eu-repo/semantics/openAccess
dc.rights.accesoAbierto (Texto Completo)spa
dc.source.instnameinstname:Universidad del Rosariospa
dc.source.reponamereponame:Repositorio Institucional EdocURspa
dc.subject.keywordCell culturespa
dc.subject.keywordCentrifugationspa
dc.subject.keywordDamage detectionspa
dc.subject.keywordEnergy dissipationspa
dc.subject.keywordHydrodynamicsspa
dc.subject.keywordMacrophagesspa
dc.subject.keywordMembranesspa
dc.subject.keywordNitric oxidespa
dc.subject.keywordStressesspa
dc.subject.keywordSuspensions (fluids)spa
dc.subject.keywordElectrical disturbancesspa
dc.subject.keywordEnergy dissipation ratespa
dc.subject.keywordHydrodynamic stressspa
dc.subject.keywordMembrane potentialsspa
dc.subject.keywordMurine macrophagesspa
dc.subject.keywordOperating conditionspa
dc.subject.keywordOptimal operating conditionsspa
dc.subject.keywordSensitive indicatorspa
dc.subject.keywordBioinformaticsspa
dc.subject.keywordFampridinespa
dc.subject.keywordNitric oxidespa
dc.subject.keywordSodium chloridespa
dc.subject.keywordAnimal cellspa
dc.subject.keywordArticlespa
dc.subject.keywordCell culturespa
dc.subject.keywordCell damagespa
dc.subject.keywordCell functionspa
dc.subject.keywordCell structurespa
dc.subject.keywordCell suspensionspa
dc.subject.keywordCell viabilityspa
dc.subject.keywordCentrifugationspa
dc.subject.keywordComparative effectivenessspa
dc.subject.keywordControlled studyspa
dc.subject.keywordCorrelation analysisspa
dc.subject.keywordFractionationspa
dc.subject.keywordHyperpolarizationspa
dc.subject.keywordIntermethod comparisonspa
dc.subject.keywordJ774.2 cell linespa
dc.subject.keywordMacrophagespa
dc.subject.keywordMembrane hyperpolarizationspa
dc.subject.keywordMembrane potentialspa
dc.subject.keywordMousespa
dc.subject.keywordNonhumanspa
dc.subject.keywordPressurespa
dc.subject.keywordPriority journalspa
dc.subject.keywordShear stressspa
dc.subject.keywordSplit flow fractionationspa
dc.subject.keywordAnimalspa
dc.subject.keywordCell linespa
dc.subject.keywordChemistryspa
dc.subject.keywordCytologyspa
dc.subject.keywordFractionationspa
dc.subject.keywordProceduresspa
dc.subject.keywordAnimalsspa
dc.subject.keywordCell Linespa
dc.subject.keywordCentrifugationspa
dc.subject.keywordChemical Fractionationspa
dc.subject.keywordMacrophagesspa
dc.subject.keywordMicespa
dc.subject.keywordCentrifugationspa
dc.subject.keywordEnergy dissipation ratespa
dc.subject.keywordHydrodynamic damagespa
dc.subject.keywordMacrophagesspa
dc.subject.keywordMembrane potentialspa
dc.subject.keywordSplit-flow fractionationspa
dc.titleMorphological and electrical disturbances after split-flow fractionation in murine macrophagesspa
dc.typearticleeng
dc.type.hasVersioninfo:eu-repo/semantics/publishedVersion
dc.type.spaArtículospa
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