Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories
Roger W L Godschalk1, Clara Ersson2, Maciej Stępnik3
1Department of Toxicology, Maastricht University, Universiteitssingel 50, Maastricht, Limburg 6200MD, the Netherlands, Department of Biosciences and Nutrition, Karolinska Institutet, Hälsovägen 7-9, Huddinge 14183, Stockholm, Sweden, Toxicology and Carcinogenesis Department, Nofer Institute of Occupational Medicine, 91-348 Łódź, Poland, Environmental Health Department, National Institute of Health, Rua Alexandre Herculano 321, Porto 4000-055, Portugal, Department of Cancer Studies and Molecular Medicine, University of Leicester, Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester LE2 7LX, UK, Department of Public Health, University of Copenhagen, Section of Environmental Health, Øster Farimagsgade 5, Copenhagen K 1014, Denmark, Department of Nutrition, Faculty of Medicine, University of Oslo, PB1046 Blindern, Oslo 0316, Norway, DICOMOSA Group, Department of Psychology, Area of Psychobiology, Edificio de Servicios Centrales de Investigación, Universidade da Coruña, Campus Elviña s/n, 15071-A, Coruña, Spain, Unit of Clinical and Molecular Epidemiology, IRCCS San Raffaele Pisana, Via di Val Cannuta, 247, 00166, Rome, Italy, Oxidative Stress Group, Department Cancer Studies and Molecular Medicine, University of Leicester, Leicester LE2 7LX, UK, Analytical and Environmental Sciences Division, School of Biomedical Sciences, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK, Leeds Institute of Genetics, Health and Therapeutics, University of Leeds, Leeds LS2 9JT, UK, Department of Food Science and Microbiology, Division of Human Nutrition, University of Milan, via Celoria 2, Milan 20133, Italy, Department Food Sciences and Toxicology, University of Navarra, C/ Irunlarrea 1, Pamplona 31008, Spain, Department of Medical Sciences, Via Santena 19, University of Turin, Turin 10126, Italy, Human Genetics Foundation (HuGeF), Via Nizza 52,Turin 10126, Italy.
Abstract:
This study investigated the levels of DNA strand breaks and formamidopyrimidine DNA glycosylase (FPG) sensitive sites, as assessed by the comet assay, in peripheral blood mononuclear cells (PBMC) from healthy women from five different countries in Europe. The laboratory in each country (referred to as 'centre') collected and cryopreserved PBMC samples from three donors, using a standardised cell isolation protocol. The samples were analysed in 13 different laboratories for DNA damage, which is measured by the comet assay. The study aim was to assess variation in DNA damage in PBMC samples that were collected in the same way and processed using the same blood isolation procedure. The inter-laboratory variation was the prominent contributor to the overall variation. The inter-laboratory coefficient of variation decreased for both DNA strand breaks (from 68 to 26%) and FPG sensitive sites (from 57 to 12%) by standardisation of the primary comet assay endpoint with calibration curve samples. The level of DNA strand breaks in the samples from two of the centres (0.56-0.61 lesions/10(6) bp) was significantly higher compared with the other three centres (0.41-0.45 lesions/10(6) bp). In contrast, there was no difference between the levels of FPG sensitive sites in PBMC samples from healthy donors in the different centres (0.41-0.52 lesion/10(6) bp).
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