Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories

Roger W L Godschalk; Clara Ersson; Maciej Stępnik; Magdalena Ferlińska; Jadwiga Palus; João Paulo Teixeira; Solange Costa; George D D Jones; Jennifer A Higgins; Johanna Kain; Lennart Möller; Lykke Forchhammer; Steffen Loft; Yolanda Lorenzo; Andrew R Collins; Frederik-Jan van Schooten; Blanca Laffon; Vanessa Valdiglesias; Marcus Cooke; Vilas Mistry; Mahsa Karbaschi; David H Phillips; Osman Sozeri; Michael N Routledge; Kirsty Nelson-Smith; Patrizia Riso; Marisa Porrini; Adela López de Cerain; Amaya Azqueta; Giuseppe Matullo; Alessandra Allione; Peter Møller

doi:10.1093/mutage/geu012

Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories

Roger W L Godschalk, Clara Ersson, Maciej Stępnik, Magdalena Ferlińska, Jadwiga Palus, João Paulo Teixeira, Solange Costa, George D D Jones, Jennifer A Higgins, Johanna Kain, Lennart Möller, Lykke Forchhammer, Steffen Loft, Yolanda Lorenzo, Andrew R Collins, Frederik-Jan van Schooten, Blanca Laffon, Vanessa Valdiglesias, Marcus Cooke, Vilas MistryMahsa Karbaschi, David H Phillips, Osman Sozeri, Michael N Routledge, Kirsty Nelson-Smith, Patrizia Riso, Marisa Porrini, Adela López de Cerain, Amaya Azqueta, Giuseppe Matullo, Alessandra Allione, Peter Møller

18 Citations (Scopus)

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).

Original language	English
Journal	Mutagenesis
Volume	29
Issue number	4
Pages (from-to)	241-9
Number of pages	9
ISSN	0267-8357
DOIs	https://doi.org/10.1093/mutage/geu012
Publication status	Published - 1 Jul 2014

Keywords

Adult
Calibration
Cell Separation
Comet Assay
DNA Breaks, Double-Stranded
DNA Damage
DNA-Formamidopyrimidine Glycosylase
Female
Humans
Laboratories
Leukocytes, Mononuclear
Middle Aged
Mutagenicity Tests
Regression Analysis

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Godschalk, R. W. L., Ersson, C., Stępnik, M., Ferlińska, M., Palus, J., Teixeira, J. P., Costa, S., Jones, G. D. D., Higgins, J. A., Kain, J., Möller, L., Forchhammer, L., Loft, S., Lorenzo, Y., Collins, A. R., van Schooten, F-J., Laffon, B., Valdiglesias, V., Cooke, M., ... Møller, P. (2014). Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories. Mutagenesis, 29(4), 241-9. https://doi.org/10.1093/mutage/geu012

Godschalk, RWL, Ersson, C, Stępnik, M, Ferlińska, M, Palus, J, Teixeira, JP, Costa, S, Jones, GDD, Higgins, JA, Kain, J, Möller, L, Forchhammer, L, Loft, S, Lorenzo, Y, Collins, AR, van Schooten, F-J, Laffon, B, Valdiglesias, V, Cooke, M, Mistry, V, Karbaschi, M, Phillips, DH, Sozeri, O, Routledge, MN, Nelson-Smith, K, Riso, P, Porrini, M, López de Cerain, A, Azqueta, A, Matullo, G, Allione, A & Møller, P 2014, 'Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories', Mutagenesis, vol. 29, no. 4, pp. 241-9. https://doi.org/10.1093/mutage/geu012

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title = "Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories",

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).",

keywords = "Adult, Calibration, Cell Separation, Comet Assay, DNA Breaks, Double-Stranded, DNA Damage, DNA-Formamidopyrimidine Glycosylase, Female, Humans, Laboratories, Leukocytes, Mononuclear, Middle Aged, Mutagenicity Tests, Regression Analysis",

author = "Godschalk, {Roger W L} and Clara Ersson and Maciej St{\c e}pnik and Magdalena Ferli{\'n}ska and Jadwiga Palus and Teixeira, {Jo{\~a}o Paulo} and Solange Costa and Jones, {George D D} and Higgins, {Jennifer A} and Johanna Kain and Lennart M{\"o}ller and Lykke Forchhammer and Steffen Loft and Yolanda Lorenzo and Collins, {Andrew R} and {van Schooten}, Frederik-Jan and Blanca Laffon and Vanessa Valdiglesias and Marcus Cooke and Vilas Mistry and Mahsa Karbaschi and Phillips, {David H} and Osman Sozeri and Routledge, {Michael N} and Kirsty Nelson-Smith and Patrizia Riso and Marisa Porrini and {L{\'o}pez de Cerain}, Adela and Amaya Azqueta and Giuseppe Matullo and Alessandra Allione and Peter M{\o}ller",

note = "{\textcopyright} The Author 2014. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: [email protected].",

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doi = "10.1093/mutage/geu012",

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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

PY - 2014/7/1

Y1 - 2014/7/1

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

SN - 0267-8357

VL - 29

SP - 241

EP - 249

JO - Mutagenesis

JF - Mutagenesis

IS - 4

ER -

Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories

Abstract

Keywords

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