title: Epigenetic Classification of Human Mesenchymal Stromal Cells.
abstract: Standardization of mesenchymal stromal cells (MSCs) is hampered by the lack of a precise definition for these cell preparations; for example, there are no molecular markers to discern MSCs and fibroblasts. In this study, we followed the hypothesis that specific DNA methylation (DNAm) patterns can assist classification of MSCs. We utilized 190 DNAm profiles to address the impact of tissue of origin, donor age, replicative senescence, and serum supplements on the epigenetic makeup. Based on this, we elaborated a simple epigenetic signature based on two CpG sites to classify MSCs and fibroblasts, referred to as the Epi-MSC-Score. Another two-CpG signature can distinguish between MSCs from bone marrow and adipose tissue, referred to as the Epi-Tissue-Score. These assays were validated by site-specific pyrosequencing analysis in 34 primary cell preparations. Furthermore, even individual subclones of MSCs were correctly classified by our epigenetic signatures. In summary, we propose an alternative concept to use DNAm patterns for molecular definition of cell preparations, and our epigenetic scores facilitate robust and cost-effective quality control of MSC cultures.
authors: Division of Stem Cell Biology and Cellular Engineering, Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Pauwelsstraße 20, 52074 Aachen, Germany; Department of Cell Biology, Institute for Biomedical Engineering, RWTH Aachen University Medical School, 52074 Aachen, Germany; Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-000, Brazil.
authors: Danilo Candido
authors: DC
authors: de Almeida
authors: Department of Cell Biology, IZKF Research Group Bioinformatics, Institute for Biomedical Engineering, RWTH Aachen University Medical School, 52074 Aachen, Germany; Department of Statistics, Centre for Natural and Exact Sciences, Federal University of Paraiba, João Pessoa 58051-900, Brazil.
authors: Marcelo R P
authors: MR
authors: Ferreira
authors: Division of Stem Cell Biology and Cellular Engineering, Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Pauwelsstraße 20, 52074 Aachen, Germany; Department of Cell Biology, Institute for Biomedical Engineering, RWTH Aachen University Medical School, 52074 Aachen, Germany.
authors: Julia
authors: J
authors: Franzen
authors: Division of Stem Cell Biology and Cellular Engineering, Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Pauwelsstraße 20, 52074 Aachen, Germany; Department of Cell Biology, Institute for Biomedical Engineering, RWTH Aachen University Medical School, 52074 Aachen, Germany.
authors: Carola I
authors: CI
authors: Weidner
authors: Division of Stem Cell Biology and Cellular Engineering, Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Pauwelsstraße 20, 52074 Aachen, Germany; Department of Cell Biology, Institute for Biomedical Engineering, RWTH Aachen University Medical School, 52074 Aachen, Germany.
authors: Joana
authors: J
authors: Frobel
authors: Department of Cell Biology, Institute for Biomedical Engineering, RWTH Aachen University Medical School, 52074 Aachen, Germany; Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, 52074 Aachen, Germany.
authors: Martin
authors: M
authors: Zenke
authors: Department of Cell Biology, IZKF Research Group Bioinformatics, Institute for Biomedical Engineering, RWTH Aachen University Medical School, 52074 Aachen, Germany.
authors: Ivan G
authors: IG
authors: Costa
authors: Division of Stem Cell Biology and Cellular Engineering, Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Pauwelsstraße 20, 52074 Aachen, Germany; Department of Cell Biology, Institute for Biomedical Engineering, RWTH Aachen University Medical School, 52074 Aachen, Germany. Electronic address: wwagner@ukaachen.de.
authors: Wolfgang
authors: W
authors: Wagner
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D001854:Bone Marrow Cells / Q000166:cytology; D002999:Clone Cells; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation / Q000235:genetics; D044127:Epigenesis, Genetic; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D059630:Mesenchymal Stem Cells / Q000145:classification* / Q000166:cytology / Q000378:metabolism*
keywords: DNA methylation; adipose tissue; bone marrow; epigenetic score; fibroblast; mesenchymal stem cell; mesenchymal stromal cell