Scientific publications
Pancreatic differentiation of Pdx1-GFP reporter mouse induced pluripotent stem cells
Porciuncula A (1), Kumar A (2), Rodriguez S (1), Atari M (3), Araña M (4), Martin F (5), Soria B (5), Prosper F (1), Verfaillie C (6), Barajas M (7).
(1) 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.
(2) Stem Cell Institute, KU Leuven, Leuven 3000, Belgium.
(3) Regenerative Medicine Research Institute, UIC Barcelona, Barcelona, Spain.
(4) Centro de Investigación Médica Aplicada (CIMA), University of Navarra, Avda. Pio XII 55, Pamplona 31008, Spain.
(5) CABIMER, Andalusian Center for Molecular Biology and Regenerative Medicine, Seville 41092, Spain.
(6) Stem Cell Institute, KU Leuven, Leuven 3000, Belgium; Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven, Leuven 3000, Belgium.
(7) 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
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.
CITATION Differentiation. 2016 May 12. pii: S0301-4681(16)30015-9. doi: 10.1016/j.diff.2016.04.005
Our authors
