Pancreatic differentiation of Pdx1-GFP reporter mouse induced pluripotent stem cells

Angelo Porciuncula, Anujith Kumar, Saray Rodriguez, Maher Atari, Miriam Araña, Franz Martin, Bernat Soria, Felipe Prosper, Catherine Verfaillie, Miguel Barajas

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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.

Original languageEnglish
Pages (from-to)249-256
Number of pages8
JournalDifferentiation
Volume92
Issue number5
DOIs
Publication statusPublished - 01-12-2016

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All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Developmental Biology
  • Cell Biology
  • Cancer Research

Cite this

Porciuncula, A., Kumar, A., Rodriguez, S., Atari, M., Araña, M., Martin, F., Soria, B., Prosper, F., Verfaillie, C., & Barajas, M. (2016). Pancreatic differentiation of Pdx1-GFP reporter mouse induced pluripotent stem cells. Differentiation, 92(5), 249-256. https://doi.org/10.1016/j.diff.2016.04.005