title: 5-azacytidine improves the osteogenic differentiation potential of aged human adipose-derived mesenchymal stem cells by DNA demethylation.
abstract: The therapeutic value of adipose-derived mesenchymal stem cells (Ad-MSCs) for bone regeneration is critically discussed. A possible reason for reduced osteogenic potential may be an age-related deterioration of the Ad-MSCs. In long term in vitro culture, epigenomic changes in DNA methylation are known to cause gene silencing, affecting stem cell growth as well as the differentiation potential. In this study, we observed an age-related decline in proliferation of primary human Ad-MSCs. Decreased Nanog, Oct4 and Lin28A and increased Sox2 gene-expression was accompanied by an impaired osteogenic differentiation potential of Ad-MSCs isolated from old donors (>60 a) as compared to Ad-MSCs isolated from younger donors (<45 a). 5-hydroxymethylcytosine (5 hmC) and 5-methylcytonsine (5 mC) distribution as well as TET gene expression were evaluated to assess the evidence of active DNA demethylation. We observed a decrease of 5 hmC in Ad-MSCs from older donors. Incubation of these cells with 5-Azacytidine induced proliferation and improved the osteogenic differentiation potential in these cells. The increase in AP activity and matrix mineralization was associated with an increased presence of 5 hmC as well as with an increased TET2 and TET3 gene expression. Our data show, for the first time, a decrease of DNA hydroxymethylation in Ad-MSCs which correlates with donor-age and that treatment with 5-Azacytidine provides an approach which could be used to rejuvenate Ad-MSCs from aged donors.
authors.affiliation: Siegfried Weller Institute for Trauma Research, BG Trauma Center, Eberhard Karls University Tübingen, Tübingen, Germany.
authors.forename: Xueying
authors.initials: X
authors.lastname: Yan
authors.affiliation: Siegfried Weller Institute for Trauma Research, BG Trauma Center, Eberhard Karls University Tübingen, Tübingen, Germany.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Ehnert
authors.affiliation: Clinic of Vascular and Endovascular Surgery, Klinikum rechts der Isar, Technische Universität München, Munich, Germany.
authors.forename: Mihaela
authors.initials: M
authors.lastname: Culmes
authors.affiliation: Siegfried Weller Institute for Trauma Research, BG Trauma Center, Eberhard Karls University Tübingen, Tübingen, Germany.
authors.forename: Anastasia
authors.initials: A
authors.lastname: Bachmann
authors.affiliation: Department of Trauma Surgery, Klinikum rechts der Isar, Technische Universität München, Munich, Germany.
authors.forename: Claudine
authors.initials: C
authors.lastname: Seeliger
authors.affiliation: Department of Trauma Surgery, Klinikum rechts der Isar, Technische Universität München, Munich, Germany.
authors.forename: Lilianna
authors.initials: L
authors.lastname: Schyschka
authors.affiliation: Department of Trauma Surgery, Klinikum rechts der Isar, Technische Universität München, Munich, Germany.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Clinic for Hand-, Plastic-, Reconstructive- and Vascular Surgery, BG Trauma Center, Eberhard Karls University Tübingen, Tübingen, Germany.
authors.forename: Afshin
authors.initials: A
authors.lastname: Rahmanian-Schwarz
authors.affiliation: Siegfried Weller Institute for Trauma Research, BG Trauma Center, Eberhard Karls University Tübingen, Tübingen, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Stöckle
authors.affiliation: Center for Regenerative Medicine, the University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Paul A
authors.initials: PA
authors.lastname: De Sousa
authors.affiliation: Clinic of Vascular and Endovascular Surgery, Klinikum rechts der Isar, Technische Universität München, Munich, Germany.
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Pelisek
authors.affiliation: Siegfried Weller Institute for Trauma Research, BG Trauma Center, Eberhard Karls University Tübingen, Tübingen, Germany.
authors.forename: Andreas K
authors.initials: AK
authors.lastname: Nussler
mesh_terms: D052436:Adipose Tissue, White / Q000166:cytology; D000375:Aging; D001374:Azacitidine / Q000494:pharmacology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation / Q000187:drug effects*; D015254:DNA Modification Methylases / Q000037:antagonists & inhibitors; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D049308:Dioxygenases / Q000235:genetics / Q000378:metabolism; D044127:Epigenesis, Genetic; D015870:Gene Expression; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000502:physiology*; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D010012:Osteogenesis; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D000076182:Sp7 Transcription Factor; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; C493709:RUNX2 protein, human; D000076182:Sp7 Transcription Factor; C478163:SP7 protein, human; D014157:Transcription Factors; D015675:Osteocalcin; C578792:TET3 protein, human; D049308:Dioxygenases; C541081:TET2 protein, human; D015254:DNA Modification Methylases; D001374:Azacitidine