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Validation of reference genes for real-time PCR of cord blood mononuclear cells, differentiating endothelial progenitor cells, and mature endothelial cells

dc.rights.licenseopenen_US
dc.contributor.authorROYER, Caroline
dc.contributor.authorBEGIN, Andree-Anne Guay
dc.contributor.authorPLAWINSKI, Laurent
dc.contributor.authorLEVESQUE, Lucie
dc.contributor.authorDURRIEU, Marie-Christine
dc.contributor.authorLAROCHE, Gaetan
dc.date.accessioned2020-03-25T15:25:53Z
dc.date.available2020-03-25T15:25:53Z
dc.date.issued2018
dc.identifier.issn0014-4827en_US
dc.identifier.urihttps://oskar-bordeaux.fr/handle/20.500.12278/3944
dc.description.abstractEnIn the last ten years, endothelial progenitor cells (EPCs) have gained interest as an attractive cell population in regenerative medicine for vascular applications. This population is defined as the precursor of endothelial mature cells (ECs) through a process of differentiation. To our knowledge, no single marker can be used to discriminate them from mature ECs. To effectively study their differentiation kinetics, gene expression must be assessed. Quantitative real-time PCR (RT-qPCR) is widely used to analyze gene expression. To minimize the impact of variances from RT-qPCR, a rigorous selection of reference genes must be performed prior to any experiments due to variations in experimental conditions. In this study, CD34 + mononuclear cells were extracted from human cord blood and differentiated into EPCs after seeding for a maximum period of 21 days. To choose the best combinations of reference genes, we compared the results of EPCs, CD34 + mononuclear cells, and mature endothelial cells to ensure that the differentiation kinetics did not affect the expression of our selected reference genes. The expression levels of seven genes, namely, YWHAZ, GAPDH, HPRT1, RPLP0, UBC, B2M, and TBP were thus compared. The algorithms geNorm, NormFinder, BestKeeper, and the Comparative Delta Ct method were employed to assess the expression of each candidate gene. Overall results reveal that the expression stability of reference genes may differ depending on the statistical program used. YWHAZ, GAPDH, and UBC composed the optimal set of reference genes for the gene expression studies performed by RT-qPCR in our experimental conditions. This work can thus serve as a starting point for the selection of candidate reference genes to normalize the levels of gene expression in endothelial progenitor cell populations.
dc.language.isoENen_US
dc.subject.enVascular applications
dc.subject.enEndothelial progenitor cells
dc.subject.enRT-qPCR
dc.subject.enReference genes
dc.title.enValidation of reference genes for real-time PCR of cord blood mononuclear cells, differentiating endothelial progenitor cells, and mature endothelial cells
dc.typeArticle de revueen_US
dc.identifier.doi10.1016/j.yexcr.2018.07.001en_US
dc.subject.halChimie/Matériauxen_US
bordeaux.journalExperimental Cell Researchen_US
bordeaux.page389-398en_US
bordeaux.volume370en_US
bordeaux.hal.laboratoriesInstitut de Chimie & de Biologie des Membranes & des Nano-objets (CBMN) - UMR 5248
bordeaux.issue2en_US
bordeaux.institutionBordeaux INPen_US
bordeaux.institutionUniversité de Bordeauxen_US
bordeaux.peerReviewedouien_US
bordeaux.inpressnonen_US
hal.identifierhal-02519013
hal.version1
hal.date.transferred2020-03-25T15:26:09Z
hal.exporttrue
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