Hininger Isabelle, Chollat-Namy Alexia, Sauvaigo Sylvie, Osman Mireille, Faure H, Cadet Jean, Favier Alain, Roussel Anne-Marie
L.B.S.O, Université Joseph Fourier, Grenoble, France.
Mutat Res. 2004 Mar 14;558(1-2):75-80. doi: 10.1016/j.mrgentox.2003.11.004.
This study was undertaken with the aim to develop an optimised protocol for the evaluation of DNA damage in frozen whole blood. This was achieved by use of the single-cell gel electrophoresis (SCGE) or comet assay in its alkaline version. After collection of blood, the total blood sample was mixed with dimethyl sulfoxide (DMSO), a cryoprotectant commonly used for prevention of freezing-induced damage to living cells, and then stored at -80 degrees C. We observed no statistically significant differences in the level of DNA damage between fresh blood samples and frozen blood samples, as assessed by the comet assay. Considering the absence of effects of the freezing step, a frozen blood sample was included as a control sample in subsequent experiments. Thus the protocol was applied to blood samples of twenty healthy subjects including smokers and non-smokers. The comparative analysis indicated that the level of DNA damage was 56% higher in smokers than in non-smokers (P = 0.01). Altogether, this study strongly suggests that frozen whole blood could be utilised in association with the comet assay in human epidemiological bio-monitoring for the assessment of genetic damage in populations at risk.
本研究旨在开发一种优化方案，用于评估冷冻全血中的DNA损伤。这是通过使用单细胞凝胶电泳（SCGE）或碱性版本的彗星试验来实现的。采集血液后，将全血样本与二甲基亚砜（DMSO）混合，DMSO是一种常用于防止冷冻对活细胞造成损伤的冷冻保护剂，然后储存在-80摄氏度。通过彗星试验评估，我们观察到新鲜血液样本和冷冻血液样本之间的DNA损伤水平没有统计学上的显著差异。考虑到冷冻步骤没有影响，在后续实验中，将冷冻血液样本作为对照样本。因此，该方案应用于包括吸烟者和非吸烟者在内的20名健康受试者的血液样本。比较分析表明，吸烟者的DNA损伤水平比非吸烟者高56%（P = 0.01）。总之，本研究强烈表明，冷冻全血可与彗星试验联合用于人类流行病学生物监测，以评估高危人群的遗传损伤。
