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dc.contributor.authorCarballar, Reyes-
dc.contributor.authorCanyelles, Maria de Lluc-
dc.contributor.authorFernández, Claudia-
dc.contributor.authorMartí, Yasmina-
dc.contributor.authorBonnin, Sarah-
dc.contributor.authorCastaño Boldú, Esther-
dc.contributor.authorMontanya Mias, Eduard-
dc.contributor.authorTéllez i Besolí, Noèlia-
dc.description.abstractβ-cell proliferation is a rare event in adult pancreatic islets. To study the replication-related β-cell biology we designed a replicating β-cells sorting system for gene expression experiments. Replicating β-cells were identifed by EdU incorporation and purifed by fow cytometry. For β-cell separation islet cells were sorted by size, granularity and Newport Green fuorescence emission that was combined with emitted fuorescence for EdU-labelled replicating cells sorting. The purity of the resulting sorted populations was evaluated by insulin staining and EdU for β-cell identifcation and for replicating cells, respectively. Total RNA was isolated from purifed cell-sorted populations for gene expression analysis. Cell sorting of dispersed islet cells resulted in 96.2% purity for insulin positivity in the collected β-cell fraction and 100% efciency of the EdU-based cell separation. RNA integrity was similar between FACSsorted replicating and quiescent β-cells. Global transcriptome analysis of replicating vs quiescent β-cells showed the expected enrichment of categories related to cell division and DNA replication. Indeed, key genes in the spindle check-point were the most upregulated genes in replicating β-cells. This work provides a method that allows for the isolation of replicating β-cells, a very scarce population in adult pancreatic islets.-
dc.format.extent10 p.-
dc.publisherNature Publishing Group-
dc.relation.isformatofReproducció del document publicat a:
dc.relation.ispartofScientific Reports, 2017, vol. 7, p. 17515-
dc.rightscc-by (c) Carballar, Reyes et al., 2017-
dc.subject.classificationExpressió gènica-
dc.subject.classificationMalalties del pàncrees-
dc.subject.otherGene expression-
dc.subject.otherPancréas diseases-
dc.titlePurification of replicating pancreatic β-cells for gene expression studies-
Appears in Collections:Articles publicats en revistes (Ciències Fisiològiques)
Articles publicats en revistes (Ciències Clíniques)
Articles publicats en revistes (Institut d'lnvestigació Biomèdica de Bellvitge (IDIBELL))
Articles publicats en revistes (Centres Científics i Tecnològics de la Universitat de Barcelona (CCiTUB))

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