title: Epigenetic Induction of Definitive and Pancreatic Endoderm Cell Fate in Human Fibroblasts.
abstract: Reprogramming can occur by the introduction of key transcription factors (TFs) as well as by epigenetic changes. We demonstrated that histone deacetylase inhibitor (HDACi) Trichostatin A (TSA) combined with a chromatin remodeling medium (CRM) induced expression of a number of definitive endoderm and early and late pancreatic marker genes. When CRM was omitted, endoderm/pancreatic marker genes were not induced. Furthermore, treatment with DNA methyltransferase inhibitor (DNMTi) 5-azacytidine (5AZA) CRM did not affect gene expression changes, and when 5AZA was combined with TSA, no further increase in gene expression of endoderm, pancreatic endoderm, and endocrine markers was seen over levels induced with TSA alone. Interestingly, TSA-CRM did not affect expression of pluripotency and hepatocyte genes but induced some mesoderm transcripts. Upon removal of TSA-CRM, the endoderm/pancreatic gene expression profile returned to baseline. Our findings underscore the role epigenetic modification in transdifferentiation of one somatic cell into another. However, full reprogramming of fibroblasts to β-cells will require combination of this approach with TF overexpression and/or culture of the partially reprogrammed cells under β-cell specific conditions.
authors.affiliation: Interdepartmental Stem Cell Institute, Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven, 3000 Leuven, Belgium.
authors.forename: Rangarajan
authors.identifier: 0000-0002-2751-3143
authors.initials: R
authors.lastname: Sambathkumar
authors.affiliation: Interdepartmental Stem Cell Institute, Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven, 3000 Leuven, Belgium.
authors.forename: Eric
authors.identifier: 0000-0002-6770-3857
authors.initials: E
authors.lastname: Kalo
authors.affiliation: Interdepartmental Stem Cell Institute, Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven, 3000 Leuven, Belgium.
authors.forename: Rob
authors.identifier: 0000-0003-1046-5509
authors.initials: R
authors.lastname: Van Rossom
authors.affiliation: University Medical Center Groningen (UMCG), Pathology and Medical Biology, Section of Immunoendocrinology, University of Groningen, Hanzeplein 1, EA 11, 9713 GZ Groningen, Netherlands.
authors.forename: Marijke M
authors.initials: MM
authors.lastname: Faas
authors.affiliation: University Medical Center Groningen (UMCG), Pathology and Medical Biology, Section of Immunoendocrinology, University of Groningen, Hanzeplein 1, EA 11, 9713 GZ Groningen, Netherlands.
authors.forename: Paul
authors.initials: P
authors.lastname: de Vos
authors.affiliation: Interdepartmental Stem Cell Institute, Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven, 3000 Leuven, Belgium.
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Verfaillie