Epigenetic Induction of Definitive and Pancreatic Endoderm Cell Fate in Human Fibroblasts

Figure 1

Trichostatin A (TSA) treatment of primary adult fibroblasts induces transient definitive endoderm and pancreatic endoderm markers. (a) qRT-PCR analysis demonstrated induction of endodermal genes (GATA4, SOX7, MIXL1, EOMES, E-CADHERIN, SOX17, FOXA2, and CXCR4 but not GSC) in TSA-CRM treated primary human fibroblast. (b) qRT-PCR analysis demonstrated induction of pancreatic endoderm genes (PTF1A, PDX1, HLXB9, and NKX6.1 but not SOX9, PAX6, and NEUROD1) in TSA-CRM treated primary human fibroblast. (c) qRT-PCR analysis demonstrated induction of pancreatic endocrine genes (ISL1, ARX, and MAFB but not PAX4, NGN3, MAFA, INS, SST, and GCG) in TSA-CRM treated primary human fibroblast. (d) qRT-PCR analysis demonstrated that hepatocyte genes (ALB and HNF4A) were not induced except for AFP. (e) qRT-PCR analysis demonstrated induction of mesoderm lineage genes (MYOD1 and FLK1 but not TIE-2 and VE-CADHERIN) in TSA-CRM treated primary human fibroblast. (f) qRT-PCR analysis demonstrated no induction of pluripotency genes (OCT4, SOX2, and NANOG) expression in TSA-CRM treated primary human fibroblast. Black bar, CRM treated primary human fibroblast cells; light grey bar, DMSO-CRM primary human fibroblast treated cells; dark grey bar, 100 μM TSA-CRM treated primary human fibroblast cells. Gene expression is shown relative to the housekeeping gene GAPDH. Data represent the mean ± SEM (standard error of mean) of three independent experiments. Statistical significance tests were performed between TSA treated versus untreated fibroblast and TSA treated versus DMSO treated fibroblast. , , and by unpaired 2-tailed Student’s -test. NS, not significant.

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