• Open Access

DNA methylation pattern changes upon long-term culture and aging of human mesenchymal stromal cells

Authors

  • Simone Bork,

    1. Department of Medicine V, University of Heidelberg, 69120 Heidelberg, Germany
    2. Heidelberg Academy of Sciences and Humanities, 69117 Heidelberg, Germany
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    • *

      These authors contributed equally to this study.

  • Stefan Pfister,

    1. Heidelberg Academy of Sciences and Humanities, 69117 Heidelberg, Germany
    2. Department of Pediatric Oncology, Hematology and Immunology, University of Heidelberg, 69120 Heidelberg, Germany
    3. Molecular Genetics of Pediatric Brain Tumors, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany
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    • *

      These authors contributed equally to this study.

  • Hendrik Witt,

    1. Department of Pediatric Oncology, Hematology and Immunology, University of Heidelberg, 69120 Heidelberg, Germany
    2. Molecular Genetics of Pediatric Brain Tumors, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany
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  • Patrick Horn,

    1. Department of Medicine V, University of Heidelberg, 69120 Heidelberg, Germany
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  • Bernhard Korn,

    1. Genomics and Proteomics Core Facility, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany
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  • Anthony D. Ho,

    1. Department of Medicine V, University of Heidelberg, 69120 Heidelberg, Germany
    2. Heidelberg Academy of Sciences and Humanities, 69117 Heidelberg, Germany
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  • Wolfgang Wagner

    1. Department of Medicine V, University of Heidelberg, 69120 Heidelberg, Germany
    2. Heidelberg Academy of Sciences and Humanities, 69117 Heidelberg, Germany
    3. Helmholtz-Institute for Biomedical Engineering, Aachen University Medical School, 52074 Aachen, Germany
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Wolfgang Wagner, MD, PhD, Helmholtz-Institute for Biomedical Engineering, Aachen University Medical School, Pauwelsstrasse 20, 52074 Aachen, Germany. Tel.: +49 241 8088611;e-mail: wwagner@ukaachen.de

Summary

Within 2–3 months of in vitro culture-expansion, mesenchymal stromal cells (MSC) undergo replicative senescence characterized by cell enlargement, loss of differentiation potential and ultimate growth arrest. In this study, we have analyzed DNA methylation changes upon long-term culture of MSC by using the HumanMethylation27 BeadChip microarray assessing 27 578 unique CpG sites. Furthermore, we have compared MSC from young and elderly donors. Overall, methylation patterns were maintained throughout both long-term culture and aging but highly significant differences were observed at specific CpG sites. Many of these differences were observed in homeobox genes and genes involved in cell differentiation. Methylation changes were verified by pyrosequencing after bisulfite conversion and compared to gene expression data. Notably, methylation changes in MSC were overlapping in long-term culture and aging in vivo. This supports the notion that replicative senescence and aging represent developmental processes that are regulated by specific epigenetic modifications.

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