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...
- 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
- Rights
- License
- Abierto (Texto Completo)
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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 |
_version_ |
1814167503230730240 |