title: Pancreatic differentiation of Pdx1-GFP reporter mouse induced pluripotent stem cells.
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.
authors.affiliation: Centro de Investigación Médica Aplicada (CIMA), University of Navarra, Avda. Pio XII 55, Pamplona 31008, Spain; Hematology and Cell Therapy Area, Clínica Universidad de Navarra, Avda. Pio XII 36, 31008 Pamplona, Spain.
authors.forename: Angelo
authors.initials: A
authors.lastname: Porciuncula
authors.affiliation: Stem Cell Institute, KU Leuven, Leuven 3000, Belgium.
authors.forename: Anujith
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Centro de Investigación Médica Aplicada (CIMA), University of Navarra, Avda. Pio XII 55, Pamplona 31008, Spain; Hematology and Cell Therapy Area, Clínica Universidad de Navarra, Avda. Pio XII 36, 31008 Pamplona, Spain.
authors.forename: Saray
authors.initials: S
authors.lastname: Rodriguez
authors.affiliation: Regenerative Medicine Research Institute, UIC Barcelona, Barcelona, Spain.
authors.forename: Maher
authors.initials: M
authors.lastname: Atari
authors.affiliation: Centro de Investigación Médica Aplicada (CIMA), University of Navarra, Avda. Pio XII 55, Pamplona 31008, Spain.
authors.forename: Miriam
authors.initials: M
authors.lastname: Araña
authors.affiliation: CABIMER, Andalusian Center for Molecular Biology and Regenerative Medicine, Seville 41092, Spain.
authors.forename: Franz
authors.initials: F
authors.lastname: Martin
authors.affiliation: CABIMER, Andalusian Center for Molecular Biology and Regenerative Medicine, Seville 41092, Spain.
authors.forename: Bernat
authors.initials: B
authors.lastname: Soria
authors.affiliation: Centro de Investigación Médica Aplicada (CIMA), University of Navarra, Avda. Pio XII 55, Pamplona 31008, Spain; Hematology and Cell Therapy Area, Clínica Universidad de Navarra, Avda. Pio XII 36, 31008 Pamplona, Spain.
authors.forename: Felipe
authors.initials: F
authors.lastname: Prosper
authors.affiliation: Stem Cell Institute, KU Leuven, Leuven 3000, Belgium; Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven, Leuven 3000, Belgium.
authors.forename: Catherine
authors.initials: C
authors.lastname: Verfaillie
authors.affiliation: Centro de Investigación Médica Aplicada (CIMA), University of Navarra, Avda. Pio XII 55, Pamplona 31008, Spain; Hematology and Cell Therapy Area, Clínica Universidad de Navarra, Avda. Pio XII 36, 31008 Pamplona, Spain. Electronic address: miguel.barajas@unavarra.es.
authors.forename: Miguel
authors.initials: M
authors.lastname: Barajas
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D053595:Embryonic Stem Cells / Q000166:cytology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D051379:Mice; D008822:Mice, Transgenic; D010179:Pancreas / Q000166:cytology / Q000254:growth & development*; D011401:Promoter Regions, Genetic; D015534:Trans-Activators / Q000235:genetics*
chemical_list: D018398:Homeodomain Proteins; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D049452:Green Fluorescent Proteins
keywords: Differentiation; Induced pluripotent stem cells; Mouse; Pancreas; Pdx1; Reprogramming