Please use this identifier to cite or link to this item: http://hdl.handle.net/2445/128617
Title: Purification of replicative pancreatic β-cells for gene expression studies
Author: Carballar, Reyes
Canyelles, Maria de Lluc
Fernández, Claudia
Martí, Yasmina
Bonnin, Sarah
Castaño Boldú, Esther
Montanya Mias, Eduard
Téllez i Besolí, Noèlia
Keywords: Purificació
Cèl·lules B
Pàncrees
Proliferació cel·lular
RNA
Duplicació de l'ADN
Purification
B cells
Pancreas
Cell proliferation
RNA
DNA replication
Issue Date: 13-Dec-2017
Publisher: Nature Publishing Group
Abstract: 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.
Note: Reproducció del document publicat a: https://doi.org/10.1038/s41598-017-17776-2
It is part of: Scientific Reports, 2017, vol. 13, num. 7, p. 17515
URI: http://hdl.handle.net/2445/128617
Related resource: https://doi.org/10.1038/s41598-017-17776-2
ISSN: 2045-2322
Appears in Collections:Articles publicats en revistes (Centres Científics i Tecnològics de la Universitat de Barcelona (CCiTUB))
Articles publicats en revistes (Ciències Clíniques)

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