Noticias29.05.2018Publicación del artículo “Purification of replicating pancreatic β-cells for gene expression studies” |
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La Dra. Esther Castaño, Responsable de la tecnología de Citometría y Genómica de los CCiTUB del Campus Bellvitge, ha publicado el artículo: “Purification of replicating pancreatic β-cells for gene expression studies” en la revista Molecular Pharmaceutics publicada en diciembre. En este trabajo se ha utilizado la tecnología de citometría y genómica de los Centros Científicos y Tecnológicos de la UB. Los autores de este artículo (Noelia Téllez, Reyes Carballar y otros investigadores) han usado los servicios de la unidad de Biología de Bellvitge de los CCiTUB básicamente en tres aplicaciones: - La identificación por citometría de las células beta-pancreáticas replicantes y su purificación (sorting). - El análisis de la integridad del RNA extraido de estas células - El análisis de la expresión génica de las células purificadas mediante RT-PCR El resumen del artículo es el siguiente: "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." Para más información, pueden acceder al siguiente link:[+]. |
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