Hacettepe University Faculty of Pharmacy Department of Pharmaceutical Toxicology, Ankara, Turkey.
University of Health Sciences Turkey Gülhane Faculty of Pharmacy Department of Pharmaceutical Toxicology, Ankara, Turkey.
Food Chem Toxicol. 2021 Jun;152:112163. doi: 10.1016/j.fct.2021.112163. Epub 2021 Apr 6.
Comet assay, applied to in vitro, in vivo and ex vivo systems, is a quick, simple, and sensitive method for the detection of genotoxicity. In general, fresh whole blood or peripheral blood mononuclear cells (PBMCs) are used in the assay for the determination of DNA damage and repair. In this study, the effects of storage conditions on genotoxicity assessed by Comet assay in human whole blood and lymphocyte samples, were evaluated. Whole blood and lymphocyte samples were stored at 4 °C for 1, 2, 3, 4, 5 and 7 days; at -20 °C for 1 month and at -80 °C for 3, 6 and 12 months. 1% DMSO was used as cryoprotectant. No significant differences in DNA damage were demonstrated in all of the storage conditions and durations, and the results were similar according to the median values (p < 0.05). According to Spearman or Pearson correlations, an important correlation was found between the DNA damage of the fresh samples and the samples which were kept at -80 °C for 6 months with temperature and time (p < 0.01 for Pearson and p < 0.05 for Spearman). The results of this study indicated that blood and lymphocyte samples stored in +4 °C, -20 °C and -80 °C up to 12 months can be used instead of fresh samples especially in human biomonitoring studies.
彗星试验，应用于体外、体内和离体系统，是一种快速、简单、灵敏的检测遗传毒性的方法。通常，在测定 DNA 损伤和修复时，该试验使用新鲜的全血或外周血单个核细胞（PBMCs）。在这项研究中，评估了储存条件对人全血和淋巴细胞样本彗星试验评估的遗传毒性的影响。全血和淋巴细胞样本在 4°C 下储存 1、2、3、4、5 和 7 天；在-20°C 下储存 1 个月，在-80°C 下储存 3、6 和 12 个月。1%DMSO 用作冷冻保护剂。在所有储存条件和时间下，DNA 损伤均无显著差异，中位数（p<0.05）结果相似。根据 Spearman 或 Pearson 相关性，在新鲜样本与在-80°C 下储存 6 个月的样本之间发现了重要的相关性，与温度和时间相关（Pearson 相关系数为 p<0.01，Spearman 相关系数为 p<0.05）。这项研究的结果表明，在+4°C、-20°C 和-80°C 下储存长达 12 个月的血液和淋巴细胞样本可以代替新鲜样本，特别是在人类生物监测研究中。
