TY - JOUR
T1 - Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies
AU - Møller, Peter
AU - Bankoglu, Ezgi Eyluel
AU - Stopper, Helga
AU - Giovannelli, Lisa
AU - Ladeira, Carina
AU - Koppen, Gudrun
AU - Gajski, Goran
AU - Collins, Andrew
AU - Valdiglesias, Vanessa
AU - Laffon, Blanca
AU - Boutet-Robinet, Elisa
AU - Perdry, Herve
AU - Del Bo', Cristian
AU - Langie, Sabine A. S.
AU - Dusinska, Maria
AU - Azqueta, Amaya
PY - 2021
Y1 - 2021
N2 - 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.
AB - 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.
KW - BREAST-CANCER PATIENTS
KW - CRYOPRESERVED LYMPHOCYTES
KW - PERIPHERAL LYMPHOCYTES
KW - GEL-ELECTROPHORESIS
KW - SEASONAL-VARIATIONS
KW - OXIDATIVE DAMAGE
KW - MAMMALIAN-CELLS
KW - WHOLE-BLOOD
KW - IN-VITRO
KW - VALIDATION
U2 - 10.1093/mutage/geab012
DO - 10.1093/mutage/geab012
M3 - Review
C2 - 33755160
VL - 36
SP - 193
EP - 212
JO - Mutagenesis
JF - Mutagenesis
SN - 0267-8357
IS - 3
ER -