Centres Científics i Tecnològics UB

Notícies

29.05.2018

Publicació de l'article “Purification of replicating pancreatic β-cells for gene expression studies”
La Dra. Esther Castaño, Responsable de la tecnologia de Citometria i Genòmica dels CCiTUB al Campus de Bellvitge, ha publicat l'article: “Purification of replicating pancreatic β-cells for gene expression studies” a la revista Scientific Reports publicada el passat desembre. En aquest treball s’ha utilitzat la tecnologia de citometria i genòmica dels Centres Científics i Tecnològics de la UB.

Els autors d'aquest article (Noelia Téllez, Reyes Carballar i altres investigadors) han usat els serveis de la unitat de Biologia de Bellvitge dels CCiTUB bàsicament en tres aplicacions:
- La identificació per citometria de les cèl·lules beta-pancreàtiques replicants i la seva purificació (sorting).
- L’anàlisi de la integritat del RNA extret d’aquestes cèl·lules
- L’anàlisi de la expressió gènica de les cèl·lules purificades mitjançant RT-PCR

El resum de l'article és el següent:

"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."

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