Short- and long-term reproducibility of the COMET assay for measuring DNA damage biomarkers in frozen blood samples of the EPIC-Heidelberg cohort
Autor: | Ezgi Eyluel Bankoglu, Trasias Mukama, Verena Katzke, Franzisca Stipp, Theron Johnson, Tilman Kühn, Florian Seyfried, Roger Godschalk, Andrew Collins, Rudolf Kaaks, Helga Stopper |
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Přispěvatelé: | Farmacologie en Toxicologie, RS: NUTRIM - R3 - Respiratory & Age-related Health |
Rok vydání: | 2021 |
Předmět: |
Health
Toxicology and Mutagenesis Mononuclear FREQUENCY SDG 3 - Good Health and Well-being INTER-LABORATORY VARIATION Genetics Leukocytes Humans Obesity Prospective Studies Biobank Retrospective Studies RISK Cryopreservation MICRONUCLEI Morbid/blood Reproducibility of Results Obesity Morbid/blood CANCER HUMAN-LYMPHOCYTES Obesity Morbid Human biomonitoring BODY-MASS INDEX Leukocytes Mononuclear Comet Assay SMOKING EPIC Biomarkers DNA Damage |
Zdroj: | Bankoglu, E E, Mukama, T, Katzke, V, Stipp, F, Johnson, T, Kühn, T, Seyfried, F, Godschalk, R, Collins, A, Kaaks, R & Stopper, H 2022, ' Short-and long-term reproducibility of the COMET assay for measuring DNA damage biomarkers in frozen blood samples of the EPIC-Heidelberg cohort ', Mutation Research/Genetic Toxicology and Environmental Mutagenesis, vol. 874-875, 503442 . https://doi.org/10.1016/j.mrgentox.2022.503442 Mutation Research-Genetic Toxicology and Environmental Mutagenesis, 874-875:503442. Elsevier |
ISSN: | 1879-3592 1383-5718 |
Popis: | The comet assay is widely used for quantification of genomic damage in humans. Peripheral blood derived mononuclear cells (PBMCs) are the most often used cell type for this purpose. Since the comet assay can be performed in an enhanced throughput format, it can be applied to large sample collections such as biobanks. The European Prospective Investigation into Cancer and Nutrition (EPIC) study is one of the largest existing prospective cohort studies, and the German Cancer Research Institute (DKFZ) in Heidelberg is a participating center with 25.000 frozen blood samples stored from around 25 years ago, enabling retrospective assessment of disease risk factors. However, experience with decades long frozen samples in the comet assay is so far missing. In Heidelberg, 800 study participants were re-invited twice between 2010 and 2012 to donate further blood samples. Here, we analyzed 299 Heidelberg-EPIC samples, compiled from frozen PBMC and buffy coat preparations selected from the different sampling time points. In addition, 47 frozen PBMC samples from morbidly obese individuals were included. For buffy coat samples, we observed a poor correlation between DNA damage in the same donors assessed at two sampling time points. Additionally, no correlation between DNA damage in buffy coat samples and PBMCs was found. For PBMCs, a good correlation was observed between samples of the same donors at the two time points. DNA damage was not affected by age and smoking status, but high BMI (>30; obesity) was associated with increased DNA damage in PBMCs. There was no indication for a threshold of a certain BMI for increased DNA damage. In conclusion, while 25 year-long stored buffy coat preparations may require adaptation of certain experimental parameters such as cell density and electrophoresis conditions, frozen PBMC biobank samples can be analyzed in the comet assay even after a decade of storage. |
Databáze: | OpenAIRE |
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