Johnson Adiv A, Naaldijk Yahaira, Hohaus Christian, Meisel Hans Jörg, Krystel Ilona, Stolzing Alexandra
Department of Ophthalmology, Mayo Clinic, Rochester, MN 55905, USA.
Translational Centre for Regenerative Medicine (TRM), Leipzig University, Leipzig, Germany.
Aging (Albany NY). 2016 Sep 15;9(2):340-352. doi: 10.18632/aging.101035.
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 HO or UV light. Tail lengths in HO-treated ADSCs were substantially higher than the tail lengths in UV-treated ADSCs. After 30 minutes of treatment with HO, 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.
脂肪来源的间充质干细胞（ADSCs）是具有多能性的干细胞，可促进治疗效果，常用于自体应用。关于ADSCs如何响应基因毒性应激以及供体年龄是否会影响DNA损伤和修复，目前知之甚少。在本研究中，我们使用彗星试验评估了来自年轻（20 - 40岁）、中年（41 - 60岁）和老年（61岁以上）供体的人ADSCs在经羟基脲（HO）或紫外线（UV）处理后的DNA损伤和修复情况。HO处理的ADSCs的尾巴长度显著高于UV处理的ADSCs的尾巴长度。在用HO处理30分钟后，用α-苯基叔丁基硝酮（PBN）或抗坏血酸（AA）预处理的ADSCs的尾巴DNA百分比显著降低。在各个时间点，用PBN或AA处理的大多数ADSCs显示出低橄榄尾矩。总体而言，作为DNA修复的指标，尾巴长度百分比和尾巴DNA百分比在30分钟时达到峰值，然后在2小时和4小时时间点降至接近对照水平。在大多数这些实验中，不同年龄的ADSCs显示出相当水平的DNA损伤，这表明供体年龄不影响培养的ADSCs中的DNA损伤反应。
