Bedoya, Francisco J.Tejedo, JRMartin, FranzSalguero-Aranda, CarmenTapia-Limonchi, RafaelHitos, Ana BelenDiez, IreneHmadcha, AbdelkrimFraga, MarioSoria, BernatCahuana Macedo, Gladys M2017-06-292017-06-292016-10Cell Transplantation vol 25 nº 10, p 1879-18920963-689710.3727/096368916X691178http://hdl.handle.net/10433/4132Pancreatic 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 KClenAtribución-NoComercial-SinDerivadas 3.0 EspañaCognizant Communication Corporationhttp://creativecommons.org/licenses/by-nc-nd/3.0/es/Embryonic stem cells (ESCs)Nitric oxide (NO)Cell differentiationInsulin-producing cellsDiabetesjournal articleopen access