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Fuchs, R; Stelzer, I; Drees, CM; Rehnolt, C; Schraml, E; Sadjak, A; Schwinger, W.
Modification of the alkaline comet assay with human mesenchymal stem cells.
Cell Biol Int. 2012; 36(1):113-117
Doi: 10.1042/CBI20110251
Web of Science
PubMed
FullText
FullText_MUG
- Führende Autor*innen der Med Uni Graz
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Fuchs Robert
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Stelzer Ingeborg
- Co-Autor*innen der Med Uni Graz
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Rehnolt Christian
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Sadjak Anton
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Schraml Elisabeth
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Schwinger Wolfgang
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- Abstract:
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MSCs (mesenchymal stem cells) are planned foruse in regenerative medicine to offset age-dependent alterations. However, MSCs are affected by replicative senescence associated with decreasing proliferation potential, telomere shortening and DNA damage during in vitro propagation. To monitor in vitro senescence, we have assessed the integrity of DNA by the alkaline comet assay. For optimization of the comet assay we have enhanced the stability of comet slides in liquid and minimized the background noise of the method by improving adhesion of agarose gels on the comet slides and concentrating cells on a defined small area on the slides. The modifications of the slide preparation increase the overall efficiency and reproducibility of the comet assay and minimize the image capture and storage. DNA damage of human MSCs during in vitro cultivation increased with time, as assessed by the comet assay, which therefore offers a fast and easy screening tool in future efforts to minimize replicative senescence of MSCs in vitro.
© The Author(s) Journal compilation © 2012 Portland Press Limited
- Find related publications in this database (using NLM MeSH Indexing)
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Cell Aging -
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Cell Proliferation -
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Cells, Cultured -
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Comet Assay - methods
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DNA Damage -
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Humans -
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Mesenchymal Stromal Cells - cytology
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Telomere - metabolism
- Find related publications in this database (Keywords)
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comet assay
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DNA damage
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human mesenchymal stem cell (MSC)
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replicative senescence