2026-04-17T21:40:08Zhttps://recercat.cat/oai/request

oai:recercat.cat:10230/351752025-12-12T04:10:01Zcom_2072_6col_2072_452952

00925njm 22002777a 4500 dc Carballar, Reyes author Canyelles, Maria de Lluc author Fernández, Claudia author Martí, Yasmina author Bonnin, Sarah author Castaño, Esther author Montanya, Eduard author Téllez, Noèlia author 2018-07-17T07:38:15Z 2018-07-17T07:38:15Z 2017 β-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 identified by EdU incorporation and purified by flow cytometry. For β-cell separation islet cells were sorted by size, granularity and Newport Green fluorescence emission that was combined with emitted fluorescence for EdU-labelled replicating cells sorting. The purity of the resulting sorted populations was evaluated by insulin staining and EdU for β-cell identification and for replicating cells, respectively. Total RNA was isolated from purified 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% efficiency of the EdU-based cell separation. RNA integrity was similar between FACS-sorted 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. Cell growth Diabetes Purification of replicating pancreatic β-cells for gene expression studies