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Title: Pancreatic differentiation of Pdx1-GFP reporter mouse induced pluripotent stem cells.
Authors: Porciuncula, Angelo
Kumar, Anujith
Rodriguez, Saray
Atari, Maher
Araña, Miriam
Martin, Franz
Soria, Bernat
Prosper, Felipe
Verfaillie, Catherine
Barajas, Miguel
Keywords: Differentiation;Induced pluripotent stem cells;Mouse;Pancreas;Pdx1;Reprogramming
metadata.dc.subject.mesh: Animals
Cell Differentiation
Embryonic Stem Cells
Genes, Reporter
Green Fluorescent Proteins
Homeodomain Proteins
Induced Pluripotent Stem Cells
Mice, Transgenic
Promoter Regions, Genetic
Issue Date: 12-May-2016
Abstract: Efficient induction of defined lineages in pluripotent stem cells constitutes the determinant step for the generation of therapeutically relevant replacement cells to potentially treat a wide range of diseases, including diabetes. Pancreatic differentiation has remained an important challenge in large part because of the need to differentiate uncommitted pluripotent stem cells into highly specialized hormone-secreting cells, which has been shown to require a developmentally informed step-by-step induction procedure. Here, in the framework of using induced pluripotent stem cells (iPSCs) to generate pancreatic cells for pancreatic diseases, we have generated and characterized iPSCs from Pdx1-GFP transgenic mice. The use of a GFP reporter knocked into the endogenous Pdx1 promoter allowed us to monitor pancreatic induction based on the expression of Pdx1, a pancreatic master transcription factor, and to isolate a pure Pdx1-GFP+ population for downstream applications. Differentiated cultures timely expressed markers specific to each stage and end-stage progenies acquired a rather immature beta-cell phenotype, characterized by polyhormonal expression even among cells highly expressing the Pdx1-GFP reporter. Our findings highlight the utility of employing a fluorescent protein reporter under the control of a master developmental gene in order to devise novel differentiation protocols for relevant cell types for degenerative diseases such as pancreatic beta cells for diabetes.
metadata.dc.identifier.doi: 10.1016/j.diff.2016.04.005
Appears in Collections:Producción 2020

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