1. Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies
- Author
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Goran Gajski, Blanca Laffon, Vanessa Valdiglesias, Peter Møller, Carina Ladeira, Amaya Azqueta, Elisa Boutet-Robinet, Gudrun Koppen, Hervé Perdry, Helga Stopper, Cristian Del Bo, Andrew Collins, Lisa Giovannelli, Maria Dusinska, Sabine A. S. Langie, Ezgi Eyluel Bankoglu, University of Copenhagen = Københavns Universitet (KU), University of Würzburg, NEUROFARBA Department [Firenze, Italy], Università degli Studi di Firenze = University of Florence [Firenze] (UNIFI), Escola Nacional de Saúde Pública (ENSP-NOVA), Universidade Nova de Lisboa = NOVA University Lisbon (NOVA), Flemish Institute for Technological Research (VITO), Institute for Medical Research and Occupational Health, Institute of Basic Medical Sciences [Oslo], Faculty of Medicine [Oslo], University of Oslo (UiO)-University of Oslo (UiO), Centro de Investigacións Científicas Avanzadas (CICA), Faculty of Sciences, University of A Coruña (UDC), Contaminants & Stress Cellulaire (ToxAlim-COMICS), ToxAlim (ToxAlim), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Ecole d'Ingénieurs de Purpan (INPT - EI Purpan), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Centre de recherche en épidémiologie et santé des populations (CESP), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Paul Brousse-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay, Department of Food Environmental and Nutritional Sciences, (DeFENS), University of Milan, School of Nutrition and Translational Research in Metabolism [Maastricht] (NUTRIM), Maastricht University [Maastricht], Norwegian Institute for Air Research (NILU), Universidad de Navarra [Pamplona] (UNAV), and hCOMET project (COST Action, CA 15132)
- Subjects
DNA Repair ,DNA damage ,Epidemiology ,Health, Toxicology and Mutagenesis ,[SDV]Life Sciences [q-bio] ,GEL-ELECTROPHORESIS ,Buffy coat ,Biology ,Toxicology ,Peripheral blood mononuclear cell ,Cryopreservation ,VALIDATION ,Andrology ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,MAMMALIAN-CELLS ,Genetics ,WHOLE-BLOOD ,Humans ,DNA damage, DNA repair, comet assay ,Leucocytes ,Comet assay ,Genetics (clinical) ,030304 developmental biology ,Whole blood ,0303 health sciences ,Blood Specimen Collection ,Molecular ,IN-VITRO ,DNA ,PERIPHERAL LYMPHOCYTES ,SEASONAL-VARIATIONS ,In vitro ,CRYOPRESERVED LYMPHOCYTES ,chemistry ,Blood Preservation ,030220 oncology & carcinogenesis ,OXIDATIVE DAMAGE ,Leukocytes, Mononuclear ,Comet Assay ,BREAST-CANCER PATIENTS ,[SDV.MHEP]Life Sciences [q-bio]/Human health and pathology ,DNA Damage - Abstract
DNA damage and repair activity are often assessed in blood samples from humans in different types of molecular epidemiology studies. However, it is not always feasible to analyse the samples on the day of collection without any type of storage. For instance, certain studies use repeated sampling of cells from the same subject or samples from different subjects collected at different time-points, and it is desirable to analyse all these samples in the same comet assay experiment. In addition, flawless comet assay analyses on frozen samples open up the possibility of using this technique on biobank material. In this article we discuss the use of cryopreserved peripheral blood mononuclear cells (PBMCs), buffy coat (BC) and whole blood (WB) for analysis of DNA damage and repair using the comet assay. The published literature and the authors’ experiences indicate that various types of blood samples can be cryopreserved with only a minor effect on the basal level of DNA damage. There is evidence to suggest that WB and PBMCs can be cryopreserved for several years without much effect on the level of DNA damage. However, care should be taken when cryopreserving WB and BCs. It is possible to use either fresh or frozen samples of blood cells, but results from fresh and frozen cells should not be used in the same dataset. The article outlines detailed protocols for the cryopreservation of PBMCs, BCs and WB samples.
- Published
- 2021