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Tracking of Replicative Senescence in Mesenchymal Stem Cells by Colony-Forming Unit Frequency

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Stem Cells and Aging

Part of the book series: Methods in Molecular Biology ((MIMB,volume 976))

Abstract

Long-term culture of mesenchymal stem cells (MSC) has major impact on cellular characteristics and differentiation potential. Numerous clinical trials raise high hopes in regenerative medicine and this necessitates reliable quality control of the cellular products—also with regard to replicative senescence. The maximum number of population doublings before entering the senescent state depends on the cell type, tissue of origin, culture medium as well as cell culture methods. Therefore, it would be valuable to predict the remaining proliferative potential in the course of culture expansion. Here, we describe a refined fibroblastic colony forming unit (CFU-f) assay which can be performed at any passage during culture expansion with simple cell culture techniques. This method is based on limiting dilutions in the 96-well format to determine the proportion of highly proliferative and clonogenic cells. The number of CFU-f declines rapidly during culture expansion. Especially at higher passages the CFU-f frequency correlates very well with the remaining cumulative population doublings. This approach can be used as quality measure to estimate the remaining proliferative potential of MSC in culture.

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Acknowledgement

This work was supported by the excellence initiative of the German federal and state governments within the START-Program of the Faculty of Medicine, RWTH Aachen, by the Stem Cell Network North Rhine Westphalia, and by the Else-Kröner Fresenius Stiftung.

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Authors and Affiliations

  1. Stem Cell Biology and Cellular Engineering, Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, Germany

    Anne Schellenberg, Hatim Hemeda & Wolfgang Wagner

Authors
  1. Anne Schellenberg
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  2. Hatim Hemeda
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  3. Wolfgang Wagner
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Corresponding author

Correspondence to Wolfgang Wagner .

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Editors and Affiliations

  1. Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Smyth Road 501, Ottawa, K1Y 8L6, Canada

    Kursad Turksen

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Schellenberg, A., Hemeda, H., Wagner, W. (2013). Tracking of Replicative Senescence in Mesenchymal Stem Cells by Colony-Forming Unit Frequency. In: Turksen, K. (eds) Stem Cells and Aging. Methods in Molecular Biology, vol 976. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-317-6_11

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  • DOI: https://doi.org/10.1007/978-1-62703-317-6_11

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-316-9

  • Online ISBN: 978-1-62703-317-6

  • eBook Packages: Springer Protocols

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