Research Paper|Volume 9, Issue 2|pp 340—352

Protective effects of alpha phenyl-tert-butyl nitrone and ascorbic acid in human adipose derived mesenchymal stem cells from differently aged donors

Adiv A. Johnson¹, Yahaira Naaldijk², Christian Hohaus³, Hans Jörg Meisel³, Ilona Krystel⁵, Alexandra Stolzing^2,⁴

¹Department of Ophthalmology, Mayo Clinic, Rochester, MN 55905, USA
²Translational Centre for Regenerative Medicine (TRM), Leipzig University, Leipzig, Germany
³Department of Neurosurgery, BG Clinic Bergmannstrost Halle, Germany
⁴Loughborough University, Centre for Biological Engineering, Wolfson School, Loughborough, UK
⁵Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany

* * Equal contribution

Received: July 1, 2016Accepted: August 30, 2016Published: September 15, 2016

Copyright: © 2016 Johnson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Adipose-derived mesenchymal stem cells (ADSCs) are multipotent stem cells that promote therapeutic effects and are frequently used in autologous applications. Little is known about how ADSCs respond to genotoxic stress and whether or not donor age affects DNA damage and repair. In this study, we used the comet assay to assess DNA damage and repair in human ADSCs derived from young (20-40 years), middle-aged (41-60 years), and older (61+ years) donors following treatment with H₂O₂ or UV light. Tail lengths in H₂O₂-treated ADSCs were substantially higher than the tail lengths in UV-treated ADSCs. After 30 minutes of treatment with H₂O₂, ADSCs preconditioned with alpha phenyl-tert-butyl nitrone (PBN) or ascorbic acid (AA) showed a significant reduction in % tail DNA. The majority of ADSCs treated with PBN or AA displayed low olive tail movements at various timepoints. In general and indicative of DNA repair, % tail length and % tail DNA peaked at 30 minutes and then decreased to near-control levels at the 2 hour and 4 hour timepoints. Differently aged ADSCs displayed comparable levels of DNA damage in the majority of these experiments, suggesting that the age of the donor does not affect the DNA damage response in cultured ADSCs.