Carballar, ReyesCanyelles, Maria de LlucFernández, ClaudiaMartí, YasminaBonnin, SarahCastaño Boldú, EstherMontanya Mias, EduardTéllez i Besolí, Noèlia2018-07-172018-07-172017-12-132045-2322https://hdl.handle.net/2445/123700β-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.10 p.application/pdfengcc-by (c) Carballar, Reyes et al., 2017http://creativecommons.org/licenses/by/3.0/esExpressió gènicaMalalties del pàncreesDiabetisGene expressionPancréas diseasesDiabetesinfo:eu-repo/semantics/article6755762018-07-17info:eu-repo/semantics/openAccess29235543