DNA-repair measurements by use of the modified comet assay: an inter-laboratory comparison within the European Comet Assay Validation Group (ECVAG)

TY - JOUR

T1 - DNA-repair measurements by use of the modified comet assay

T2 - an inter-laboratory comparison within the European Comet Assay Validation Group (ECVAG)

AU - Godschalk, Roger W L

AU - Ersson, Clara

AU - Riso, Patrizia

AU - Porrini, Marisa

AU - Langie, Sabine A S

AU - van Schooten, Frederik-Jan

AU - Azqueta, Amaya

AU - Collins, Andrew R

AU - Jones, George D D

AU - Kwok, Rachel W L

AU - Phillips, David H

AU - Sozeri, Osman

AU - Allione, Alessandra

AU - Matullo, Giuseppe

AU - Möller, Lennart

AU - Forchhammer, Lykke

AU - Loft, Steffen

AU - Møller, Peter

N1 - Copyright © 2013 Elsevier B.V. All rights reserved.

PY - 2013/9/1

Y1 - 2013/9/1

N2 - The measurement of DNA-repair activity by extracts from cells or tissues by means of the single-cell gel electrophoresis (comet) assay has a high potential to become widely used in biomonitoring studies. We assessed the inter-laboratory variation in reported values of DNA-repair activity on substrate cells that had been incubated with Ro19-8022 plus light to generate oxidatively damaged DNA. Eight laboratories assessed the DNA-repair activity of three cell lines (i.e. one epithelial and two fibroblast cell lines), starting with cell pellets or with cell extracts provided by the coordinating laboratory. There was a large inter-laboratory variation, as evidenced by the range in the mean level of repair incisions between the laboratory with the lowest (0.002incisions/10(6)bp) and highest (0.988incisions/10(6)bp) incision activity. Nevertheless, six out of eight laboratories reported the same cell line as having the highest level of DNA-repair activity. The two laboratories that reported discordant results (with another cell line having the highest level of DNA-repair activity) were those that reported to have little experience with the modified comet assay to assess DNA repair. The laboratories were also less consistent in ordering the repair activity of the other two cell lines, probably because the DNA-repair activity by extracts from these cell lines were very similar (on average approximately 60-65% of the cell line with the highest repair capacity). A significant correlation was observed between the repair activity found in the provided and the self-made cell extracts (r=0.71, P

AB - The measurement of DNA-repair activity by extracts from cells or tissues by means of the single-cell gel electrophoresis (comet) assay has a high potential to become widely used in biomonitoring studies. We assessed the inter-laboratory variation in reported values of DNA-repair activity on substrate cells that had been incubated with Ro19-8022 plus light to generate oxidatively damaged DNA. Eight laboratories assessed the DNA-repair activity of three cell lines (i.e. one epithelial and two fibroblast cell lines), starting with cell pellets or with cell extracts provided by the coordinating laboratory. There was a large inter-laboratory variation, as evidenced by the range in the mean level of repair incisions between the laboratory with the lowest (0.002incisions/10(6)bp) and highest (0.988incisions/10(6)bp) incision activity. Nevertheless, six out of eight laboratories reported the same cell line as having the highest level of DNA-repair activity. The two laboratories that reported discordant results (with another cell line having the highest level of DNA-repair activity) were those that reported to have little experience with the modified comet assay to assess DNA repair. The laboratories were also less consistent in ordering the repair activity of the other two cell lines, probably because the DNA-repair activity by extracts from these cell lines were very similar (on average approximately 60-65% of the cell line with the highest repair capacity). A significant correlation was observed between the repair activity found in the provided and the self-made cell extracts (r=0.71, P

KW - Cell Line

KW - Comet Assay

KW - DNA Damage

KW - DNA Repair

KW - Environmental Monitoring

KW - Humans

KW - Monocytes

U2 - 10.1016/j.mrgentox.2013.06.020

DO - 10.1016/j.mrgentox.2013.06.020

M3 - Journal article

C2 - 23830929

SN - 1383-5718

VL - 757

SP - 60

EP - 67

JO - Mutation Research - Genetic Toxicology and Environmental Mutagenesis

JF - Mutation Research - Genetic Toxicology and Environmental Mutagenesis

IS - 1

ER -