dc.contributor.author | Rubio-Navarro, Alfonso | |
dc.contributor.author | Guerrero-Hue, Melania | |
dc.contributor.author | Martín-Fernandez, Beatriz | |
dc.contributor.author | Cortegano, Isabel | |
dc.contributor.author | Olivares-Álvaro, Elena | |
dc.contributor.author | de Las Heras, Natalia | |
dc.contributor.author | Alia, Mario | |
dc.contributor.author | Andres, Belen de | |
dc.contributor.author | Gaspar, Maria Luisa | |
dc.contributor.author | Egido, Jesús | |
dc.contributor.author | Moreno, Juan Antonio | |
dc.date.accessioned | 2020-04-30T07:00:43Z | |
dc.date.available | 2020-04-30T07:00:43Z | |
dc.date.issued | 2016 | |
dc.identifier.citation | J Vis Exp. 2016 Oct 18;(116) | es_ES |
dc.identifier.issn | 1940-087X | es_ES |
dc.identifier.uri | http://hdl.handle.net/20.500.12105/9812 | |
dc.description.abstract | There is increasing evidence suggesting the important role of inflammation and, subsequently, macrophages in the development and progression of renal disease. Macrophages are heterogeneous cells that have been implicated in kidney injury. Macrophages may be classified into two different phenotypes: classically activated macrophages (M1 macrophages), that release pro-inflammatory cytokines and promote fibrosis; and alternatively activated macrophages (M2 macrophages) that are associated with immunoregulatory and tissue-remodeling functions. These macrophage phenotypes need to be discriminated and analyzed to determine their contribution to renal injury. However, there are scarce studies reporting consistent phenotypic and functional information about macrophage subtypes in inflammatory renal disease models, especially in rats. This fact may be related to the limited macrophage markers used in rats, contrary to mice. Therefore, novel strategies are necessary to quantify and characterize the renal content of these infiltrating cells in a reliable way. This manuscript details a protocol for kidney digestion and further phenotypic and quantitative analysis of macrophages from rat kidneys by flow cytometry. Briefly, kidneys were incubated with collagenase and total macrophages were identified according to the dual presence of CD45 (leukocytes common antigen) and CD68 (PAN macrophage marker) in live cells.This was followed by surface staining of CD86 (M1 marker) and CD163 (M2 marker). Rat peritoneal macrophages were used as positive control for macrophage marker detection by flow cytometry. Our protocol resulted in low cellular mortality and allowed characterization of different intracellular and surface protein markers, thus limiting the loss of cellular integrity observed in other protocols. Moreover, this procedure allows the use of macrophages for further techniques, including cell sorting and mRNA or protein expression studies, among others. | es_ES |
dc.description.sponsorship | This work was supported by grants from FIS/FEDER (Programa Miguel Servet: CP10/00479, PI13/00802 and PI14/00883), Spanish Society of Atherosclerosis, Spanish Society of Nephrology and Fundaciòn Renal Iñigo Alvarez de Toledo (FRIAT) to Juan Antonio Moreno. FIS/FEDER funds PI14/00386 and Instituto Reina Sofìa de Investigaciòn Nefrològica to Jesús Egido. Fundaciòn Conchita Rabago to Melania Guerrero Hue. Fundaciòn Renal Iñigo Alvarez de Toledo (FRIAT) to Alfonso Rubio Navarro. | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | JoVE | es_ES |
dc.type.hasVersion | VoR | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-sa/4.0/ | * |
dc.subject.mesh | Animals | es_ES |
dc.subject.mesh | Biomarkers | es_ES |
dc.subject.mesh | Cytokines | es_ES |
dc.subject.mesh | Inflammation | es_ES |
dc.subject.mesh | Mice | es_ES |
dc.subject.mesh | Rats | es_ES |
dc.subject.mesh | Flow Cytometry | es_ES |
dc.subject.mesh | Kidney | es_ES |
dc.subject.mesh | Macrophages | es_ES |
dc.subject.mesh | Phenotype | es_ES |
dc.title | Phenotypic Characterization of Macrophages from Rat Kidney by Flow Cytometry | es_ES |
dc.type | journal article | es_ES |
dc.rights.license | Atribución-NoComercial-CompartirIgual 4.0 Internacional | * |
dc.identifier.pubmedID | 27805599 | es_ES |
dc.format.number | 116 | es_ES |
dc.identifier.doi | 10.3791/54599 | es_ES |
dc.contributor.funder | Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF) | |
dc.contributor.funder | Instituto de Salud Carlos III | |
dc.contributor.funder | Sociedad Española de Arteriosclerosis | |
dc.contributor.funder | Sociedad Española de Nefrología | |
dc.contributor.funder | Fundación Renal Íñigo Álvarez de Toledo | |
dc.contributor.funder | Instituto Reina Sofía de Investigación en Nefrología | |
dc.contributor.funder | Fundación Conchita Rábago de Jiménez Díaz | |
dc.description.peerreviewed | Sí | es_ES |
dc.identifier.e-issn | 1940-087X | es_ES |
dc.relation.publisherversion | https://doi.org/10.3791/54599 | es_ES |
dc.identifier.journal | Journal of visualized experiments : JoVE | es_ES |
dc.repisalud.centro | ISCIII::Centro Nacional de Microbiología | es_ES |
dc.repisalud.institucion | ISCIII | es_ES |
dc.relation.projectID | info:eu_repo/grantAgreement/ES/CP10/00479 | es_ES |
dc.relation.projectID | info:eu_repo/grantAgreement/ES/PI13/00802 | es_ES |
dc.relation.projectID | info:eu_repo/grantAgreement/ES/PI14/00883 | es_ES |
dc.relation.projectID | info:eu_repo/grantAgreement/ES/PI14/00386 | es_ES |
dc.rights.accessRights | open access | es_ES |