A link between the accumulation of DNA damage and loss of multi-potency of human mesenchymal stromal cells.
Bottom Line: Human mesenchymal stromal cells (hMSCs) represent an attractive cell source for clinic applications.Unfortunately, this leads to a significant decrease in their stemness.To identify the mechanism behind loss of multi-potency, hMSCs were expanded until replicative senescence and the concomitant molecular changes were characterized at regular intervals.
Affiliation: Department of Tissue Regeneration, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, Enschede, The Netherlands. email@example.comShow MeSH
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Mentions: Next, we performed Western blot analysis on hMSCs at different RPDs and analysed the abundance of proteins involved in the DNA damage response pathway, such as phospho-p53 (Ser15), p16 (c-20) and p21Waf1/Cip1 (Fig. 3A). In accordance with the previous results, we observed an activation of the DNA damage response pathway with time of culture. There was a particularly strong increase from RPD 10 to RPD 17. We suggested that DNA damage accumulation is due to impaired DNA repair mechanisms in late passage cells. We analysed the capacity of NER because it is involved in the repair of many types of DNA damage including the ones induced by reactive oxygen species associated with premature ageing phenotypes. To this end, hMSC from different RPD were seeded onto coverslips and irradiated with UV after which we measured the incorporation of 3H-thymidine. With increased PD, there was no significant change in the NER capacity (Fig. 3B).
Affiliation: Department of Tissue Regeneration, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, Enschede, The Netherlands. firstname.lastname@example.org