%0 Journal Article 
%A Bedoya, Francisco J.
%A Tejedo, JR
%A Martin, Franz
%A Salguero-Aranda, Carmen
%A Tapia-Limonchi, Rafael
%A Hitos, Ana Belen
%A Diez, Irene
%A Hmadcha, Abdelkrim
%A Fraga, Mario
%A Soria, Bernat
%A Cahuana Macedo, Gladys M
%T Differentiation of Mouse Embryonic Stem Cells toward Functional Pancreatic ß-Cell Surrogates through Epigenetic Regulation of Pdx1 by Nitric Oxide
%D 2016
%@ 0963-6897
%U http://hdl.handle.net/10433/4132
%X Pancreatic and duodenal homeobox 1 (Pdx1) is a transcription factor that regulates the embryonic development of the pancreas and the differentiation toward ß cells. Previously, we have shown that exposure of mouse embryonic stem cells (mESCs) to high concentrations of diethylenetriamine nitric oxide adduct (DETA-NO) triggers differentiation events and promotes the expression of Pdx1. Here we report evidence that Pdx1 expression is associated with release of polycomb repressive complex 2 (PRC2) and P300 from its promoter region. These events are accompanied by epigenetic changes in bivalent markers of histones trimethylated histone H3 lysine 27 (H3K27me3) and H3K4me3, site-specific changes in DNA methylation, and no change in H3 acetylation. On the basis of these findings, we developed a protocol to differentiate mESCs toward insulin-producing cells consisting of sequential exposure to DETA-NO, valproic acid, and P300 inhibitor (C646) to enhance Pdx1 expression and a final maturation step of culture in suspension to form cell aggregates. This small molecule-based protocol succeeds in obtaining cells that express pancreatic ß-cell markers such as PDX1, INS1, GCK, and GLUT2 and respond in vitro to high glucose and KCl
%K Embryonic stem cells (ESCs)
%K Nitric oxide (NO)
%K Cell differentiation
%K Insulin-producing cells
%K Diabetes
%~ GOEDOC, SUB GOETTINGEN