Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories
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Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories. / Godschalk, Roger W L; Ersson, Clara; Stępnik, Maciej; Ferlińska, Magdalena; Palus, Jadwiga; Teixeira, João Paulo; Costa, Solange; Jones, George D D; Higgins, Jennifer A; Kain, Johanna; Möller, Lennart; Forchhammer, Lykke; Loft, Steffen; Lorenzo, Yolanda; Collins, Andrew R; van Schooten, Frederik-Jan; Laffon, Blanca; Valdiglesias, Vanessa; Cooke, Marcus; Mistry, Vilas; Karbaschi, Mahsa; Phillips, David H; Sozeri, Osman; Routledge, Michael N; Nelson-Smith, Kirsty; Riso, Patrizia; Porrini, Marisa; López de Cerain, Adela; Azqueta, Amaya; Matullo, Giuseppe; Allione, Alessandra; Møller, Peter.
In: Mutagenesis, Vol. 29, No. 4, 07.2014, p. 241-9.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories
AU - Godschalk, Roger W L
AU - Ersson, Clara
AU - Stępnik, Maciej
AU - Ferlińska, Magdalena
AU - Palus, Jadwiga
AU - Teixeira, João Paulo
AU - Costa, Solange
AU - Jones, George D D
AU - Higgins, Jennifer A
AU - Kain, Johanna
AU - Möller, Lennart
AU - Forchhammer, Lykke
AU - Loft, Steffen
AU - Lorenzo, Yolanda
AU - Collins, Andrew R
AU - van Schooten, Frederik-Jan
AU - Laffon, Blanca
AU - Valdiglesias, Vanessa
AU - Cooke, Marcus
AU - Mistry, Vilas
AU - Karbaschi, Mahsa
AU - Phillips, David H
AU - Sozeri, Osman
AU - Routledge, Michael N
AU - Nelson-Smith, Kirsty
AU - Riso, Patrizia
AU - Porrini, Marisa
AU - López de Cerain, Adela
AU - Azqueta, Amaya
AU - Matullo, Giuseppe
AU - Allione, Alessandra
AU - Møller, Peter
N1 - © The Author 2014. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.
PY - 2014/7
Y1 - 2014/7
N2 - 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).
AB - 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).
KW - Adult
KW - Calibration
KW - Cell Separation
KW - Comet Assay
KW - DNA Breaks, Double-Stranded
KW - DNA Damage
KW - DNA-Formamidopyrimidine Glycosylase
KW - Female
KW - Humans
KW - Laboratories
KW - Leukocytes, Mononuclear
KW - Middle Aged
KW - Mutagenicity Tests
KW - Regression Analysis
U2 - 10.1093/mutage/geu012
DO - 10.1093/mutage/geu012
M3 - Journal article
C2 - 24737269
VL - 29
SP - 241
EP - 249
JO - Mutagenesis
JF - Mutagenesis
SN - 0267-8357
IS - 4
ER -
ID: 132418329