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

Split-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 tha...

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Autores:
Tipo de recurso:
Fecha de publicación:
2019
Institución:
Universidad del Rosario
Repositorio:
Repositorio EdocUR - U. Rosario
Idioma:
eng
OAI Identifier:
oai:repository.urosario.edu.co:10336/22287
Acceso en línea:
https://doi.org/10.1016/j.chroma.2019.01.005
https://repository.urosario.edu.co/handle/10336/22287
Palabra clave:
Cell culture
Centrifugation
Damage detection
Energy dissipation
Hydrodynamics
Macrophages
Membranes
Nitric oxide
Stresses
Suspensions (fluids)
Electrical disturbances
Energy dissipation rate
Hydrodynamic stress
Membrane potentials
Murine macrophages
Operating condition
Optimal operating conditions
Sensitive indicator
Bioinformatics
Fampridine
Nitric oxide
Sodium chloride
Animal cell
Article
Cell culture
Cell damage
Cell function
Cell structure
Cell suspension
Cell viability
Centrifugation
Comparative effectiveness
Controlled study
Correlation analysis
Fractionation
Hyperpolarization
Intermethod comparison
J774.2 cell line
Macrophage
Membrane hyperpolarization
Membrane potential
Mouse
Nonhuman
Pressure
Priority journal
Shear stress
Split flow fractionation
Animal
Cell line
Chemistry
Cytology
Fractionation
Procedures
Animals
Cell Line
Centrifugation
Chemical Fractionation
Macrophages
Mice
Centrifugation
Energy dissipation rate
Hydrodynamic damage
Macrophages
Membrane potential
Split-flow fractionation
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License
Abierto (Texto Completo)
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spelling 52542438600e93264bf-44c4-4fe1-be3e-ccd1fb77cac3e2968016-57d7-4d06-bcac-231e2bbbfa98034810fa-d863-40c8-a9e6-cf23c614273a2020-05-25T23:55:59Z2020-05-25T23:55:59Z2019Split-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.application/pdfhttps://doi.org/10.1016/j.chroma.2019.01.005219673https://repository.urosario.edu.co/handle/10336/22287engElsevier B.V.112104Journal of Chromatography AVol. 1590Journal of Chromatography A, ISSN:219673, Vol.1590,(2019); pp. 104-112https://www.scopus.com/inward/record.uri?eid=2-s2.0-85059515678&doi=10.1016%2fj.chroma.2019.01.005&partnerID=40&md5=6567e52b5af03e42e05a679c18d4806bAbierto (Texto Completo)http://purl.org/coar/access_right/c_abf2instname:Universidad del Rosarioreponame:Repositorio Institucional EdocURCell cultureCentrifugationDamage detectionEnergy dissipationHydrodynamicsMacrophagesMembranesNitric oxideStressesSuspensions (fluids)Electrical disturbancesEnergy dissipation rateHydrodynamic stressMembrane potentialsMurine macrophagesOperating conditionOptimal operating conditionsSensitive indicatorBioinformaticsFampridineNitric oxideSodium chlorideAnimal cellArticleCell cultureCell damageCell functionCell structureCell suspensionCell viabilityCentrifugationComparative effectivenessControlled studyCorrelation analysisFractionationHyperpolarizationIntermethod comparisonJ774.2 cell lineMacrophageMembrane hyperpolarizationMembrane potentialMouseNonhumanPressurePriority journalShear stressSplit flow fractionationAnimalCell lineChemistryCytologyFractionationProceduresAnimalsCell LineCentrifugationChemical FractionationMacrophagesMiceCentrifugationEnergy dissipation rateHydrodynamic damageMacrophagesMembrane potentialSplit-flow fractionationMorphological and electrical disturbances after split-flow fractionation in murine macrophagesarticleArtículohttp://purl.org/coar/version/c_970fb48d4fbd8a85http://purl.org/coar/resource_type/c_6501Urbina Bonilla, Adriana del PilarGodoy-Silva R.Hoyos M.Camacho M.10336/22287oai:repository.urosario.edu.co:10336/222872022-05-02 07:37:17.488244https://repository.urosario.edu.coRepositorio institucional EdocURedocur@urosario.edu.co
dc.title.spa.fl_str_mv Morphological and electrical disturbances after split-flow fractionation in murine macrophages
title Morphological and electrical disturbances after split-flow fractionation in murine macrophages
spellingShingle Morphological and electrical disturbances after split-flow fractionation in murine macrophages
Cell culture
Centrifugation
Damage detection
Energy dissipation
Hydrodynamics
Macrophages
Membranes
Nitric oxide
Stresses
Suspensions (fluids)
Electrical disturbances
Energy dissipation rate
Hydrodynamic stress
Membrane potentials
Murine macrophages
Operating condition
Optimal operating conditions
Sensitive indicator
Bioinformatics
Fampridine
Nitric oxide
Sodium chloride
Animal cell
Article
Cell culture
Cell damage
Cell function
Cell structure
Cell suspension
Cell viability
Centrifugation
Comparative effectiveness
Controlled study
Correlation analysis
Fractionation
Hyperpolarization
Intermethod comparison
J774.2 cell line
Macrophage
Membrane hyperpolarization
Membrane potential
Mouse
Nonhuman
Pressure
Priority journal
Shear stress
Split flow fractionation
Animal
Cell line
Chemistry
Cytology
Fractionation
Procedures
Animals
Cell Line
Centrifugation
Chemical Fractionation
Macrophages
Mice
Centrifugation
Energy dissipation rate
Hydrodynamic damage
Macrophages
Membrane potential
Split-flow fractionation
title_short Morphological and electrical disturbances after split-flow fractionation in murine macrophages
title_full Morphological and electrical disturbances after split-flow fractionation in murine macrophages
title_fullStr Morphological and electrical disturbances after split-flow fractionation in murine macrophages
title_full_unstemmed Morphological and electrical disturbances after split-flow fractionation in murine macrophages
title_sort Morphological and electrical disturbances after split-flow fractionation in murine macrophages
dc.subject.keyword.spa.fl_str_mv Cell culture
Centrifugation
Damage detection
Energy dissipation
Hydrodynamics
Macrophages
Membranes
Nitric oxide
Stresses
Suspensions (fluids)
Electrical disturbances
Energy dissipation rate
Hydrodynamic stress
Membrane potentials
Murine macrophages
Operating condition
Optimal operating conditions
Sensitive indicator
Bioinformatics
Fampridine
Nitric oxide
Sodium chloride
Animal cell
Article
Cell culture
Cell damage
Cell function
Cell structure
Cell suspension
Cell viability
Centrifugation
Comparative effectiveness
Controlled study
Correlation analysis
Fractionation
Hyperpolarization
Intermethod comparison
J774.2 cell line
Macrophage
Membrane hyperpolarization
Membrane potential
Mouse
Nonhuman
Pressure
Priority journal
Shear stress
Split flow fractionation
Animal
Cell line
Chemistry
Cytology
Fractionation
Procedures
Animals
Cell Line
Centrifugation
Chemical Fractionation
Macrophages
Mice
Centrifugation
Energy dissipation rate
Hydrodynamic damage
Macrophages
Membrane potential
Split-flow fractionation
topic Cell culture
Centrifugation
Damage detection
Energy dissipation
Hydrodynamics
Macrophages
Membranes
Nitric oxide
Stresses
Suspensions (fluids)
Electrical disturbances
Energy dissipation rate
Hydrodynamic stress
Membrane potentials
Murine macrophages
Operating condition
Optimal operating conditions
Sensitive indicator
Bioinformatics
Fampridine
Nitric oxide
Sodium chloride
Animal cell
Article
Cell culture
Cell damage
Cell function
Cell structure
Cell suspension
Cell viability
Centrifugation
Comparative effectiveness
Controlled study
Correlation analysis
Fractionation
Hyperpolarization
Intermethod comparison
J774.2 cell line
Macrophage
Membrane hyperpolarization
Membrane potential
Mouse
Nonhuman
Pressure
Priority journal
Shear stress
Split flow fractionation
Animal
Cell line
Chemistry
Cytology
Fractionation
Procedures
Animals
Cell Line
Centrifugation
Chemical Fractionation
Macrophages
Mice
Centrifugation
Energy dissipation rate
Hydrodynamic damage
Macrophages
Membrane potential
Split-flow fractionation
description Split-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.
publishDate 2019
dc.date.created.spa.fl_str_mv 2019
dc.date.accessioned.none.fl_str_mv 2020-05-25T23:55:59Z
dc.date.available.none.fl_str_mv 2020-05-25T23:55:59Z
dc.type.eng.fl_str_mv article
dc.type.coarversion.fl_str_mv http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.coar.fl_str_mv http://purl.org/coar/resource_type/c_6501
dc.type.spa.spa.fl_str_mv Artículo
dc.identifier.doi.none.fl_str_mv https://doi.org/10.1016/j.chroma.2019.01.005
dc.identifier.issn.none.fl_str_mv 219673
dc.identifier.uri.none.fl_str_mv https://repository.urosario.edu.co/handle/10336/22287
url https://doi.org/10.1016/j.chroma.2019.01.005
https://repository.urosario.edu.co/handle/10336/22287
identifier_str_mv 219673
dc.language.iso.spa.fl_str_mv eng
language eng
dc.relation.citationEndPage.none.fl_str_mv 112
dc.relation.citationStartPage.none.fl_str_mv 104
dc.relation.citationTitle.none.fl_str_mv Journal of Chromatography A
dc.relation.citationVolume.none.fl_str_mv Vol. 1590
dc.relation.ispartof.spa.fl_str_mv Journal of Chromatography A, ISSN:219673, Vol.1590,(2019); pp. 104-112
dc.relation.uri.spa.fl_str_mv https://www.scopus.com/inward/record.uri?eid=2-s2.0-85059515678&doi=10.1016%2fj.chroma.2019.01.005&partnerID=40&md5=6567e52b5af03e42e05a679c18d4806b
dc.rights.coar.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.rights.acceso.spa.fl_str_mv Abierto (Texto Completo)
rights_invalid_str_mv Abierto (Texto Completo)
http://purl.org/coar/access_right/c_abf2
dc.format.mimetype.none.fl_str_mv application/pdf
dc.publisher.spa.fl_str_mv Elsevier B.V.
institution Universidad del Rosario
dc.source.instname.spa.fl_str_mv instname:Universidad del Rosario
dc.source.reponame.spa.fl_str_mv reponame:Repositorio Institucional EdocUR
repository.name.fl_str_mv Repositorio institucional EdocUR
repository.mail.fl_str_mv edocur@urosario.edu.co
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