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2017-12-13

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cc-by (c) Carballar, Reyes et al., 2017
Si us plau utilitzeu sempre aquest identificador per citar o enllaçar aquest document: https://hdl.handle.net/2445/123700

Purification of replicating pancreatic β-cells for gene expression studies

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Recurs relacionat

https://doi.org/10.1038/s41598-017-17776-2

Resum

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 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.

Matèries

Expressió gènica, Malalties del pàncrees, Diabetis

Matèries (anglès)

Gene expression, Pancréas diseases, Diabetes

Citació

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Articles publicats en revistes (Ciències Clíniques)
Articles publicats en revistes (Centres Científics i Tecnològics de la Universitat de Barcelona (CCiTUB))
Articles publicats en revistes (Ciències Fisiològiques)
Articles publicats en revistes (Institut d'lnvestigació Biomèdica de Bellvitge (IDIBELL))

Citació

CARBALLAR, Reyes, CANYELLES, Maria de lluc, FERNÁNDEZ, Claudia, MARTÍ, Yasmina, BONNIN, Sarah, CASTAÑO BOLDÚ, Esther, MONTANYA MIAS, Eduard, TÉLLEZ I BESOLÍ, Noèlia. Purification of replicating pancreatic β-cells for gene expression studies. _Scientific Reports_. 2017. Vol. 7, núm. 17515. [consulta: 23 de gener de 2026]. ISSN: 2045-2322. [Disponible a: https://hdl.handle.net/2445/123700]

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