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DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trial

2023-10-14Journal article Restricted access
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dc.contributor.authorMøller, Peter
dc.contributor.authorAzqueta, Amaya
dc.contributor.authorRodriguez-Garraus, Adriana
dc.contributor.authorBakuradze, Tamara
dc.contributor.authorRichling, Elke
dc.contributor.authorBankoglu, Ezgi Eyluel
dc.contributor.authorStopper, Helga
dc.contributor.authorClaudino Bastos, Victoria
dc.contributor.authorLangie, Sabine A.S.
dc.contributor.authorJensen, Annie
dc.contributor.authorRistori, Sara
dc.contributor.authorScavone, Francesca
dc.contributor.authorGiovannelli, Lisa
dc.contributor.authorWojewódzka, Maria
dc.contributor.authorKruszewski, Marcin
dc.contributor.authorValdiglesias, Vanessa
dc.contributor.authorLaffon, Blanca
dc.contributor.authorCosta, Carla
dc.contributor.authorCosta, Solange
dc.contributor.authorPaulo Teixeira, João
dc.contributor.authorMarino, Mirko
dc.contributor.authorDel Bo, Cristian
dc.contributor.authorRiso, Patrizia
dc.contributor.authorZheng, Congying
dc.contributor.authorShaposhnikov, Sergey
dc.contributor.authorCollins, Andrew
dc.date.accessioned2024-02-08T10:39:53Z
dc.date.available2024-02-08T10:39:53Z
dc.date.issued2023-10-14
dc.description.abstractThe comet assay is widely used in biomonitoring studies for the analysis of DNA damage in leukocytes and peripheral blood mononuclear cells. Rather than processing blood samples directly, it can be desirable to cryopreserve whole blood or isolated cells for later analysis by the comet assay. However, this creates concern about artificial accumulation of DNA damage during cryopreservation. In this study, 10 laboratories used standardized cryopreservation and thawing procedures of monocytic (THP-1) or lymphocytic (TK6) cells. Samples were cryopreserved in small aliquots in 50% foetal bovine serum, 40% cell culture medium, and 10% dimethyl sulphoxide. Subsequently, cryopreserved samples were analysed by the standard comet assay on three occasions over a 3-year period. Levels of DNA strand breaks in THP-1 cells were increased (four laboratories), unaltered (four laboratories), or decreased (two laboratories) by long-term storage. Pooled analysis indicates only a modest positive association between storage time and levels of DNA strand breaks in THP-1 cells (0.37% Tail DNA per year, 95% confidence interval: -0.05, 0.78). In contrast, DNA strand break levels were not increased by cryopreservation in TK6 cells. There was inter-laboratory variation in levels of DNA strand breaks in THP-1 cells (SD = 3.7% Tail DNA) and TK6 reference sample cells (SD = 9.4% Tail DNA), whereas the intra-laboratory residual variation was substantially smaller (i.e. SD = 0.4%-2.2% Tail DNA in laboratories with the smallest and largest variation). In conclusion, the study shows that accumulation of DNA strand breaks in cryopreserved mononuclear blood cell lines is not a matter of concern.pt_PT
dc.description.sponsorshipCertain authors have acknowledged funding as follows: statutory funding for INTC and IRH (M.W. and M.K.); Spanish Ministry of Science and Innovation MCIN/ AEI/10.13039/501100011033 (Grants PID2020- 113788RB-I00 and PID2020-114908 GA-I00), and Xunta de Galicia (Grant ED431B 2022/16) (V.V. and B.L.); Università degli Studi di Firenze fund RICATEN21 and RICATEN22 (L.G.).pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationMutagenesis. 2023 Oct 14;38(5):273-282. doi: 10.1093/mutage/gead019pt_PT
dc.identifier.doi10.1093/mutage/gead019pt_PT
dc.identifier.issn0267-8357
dc.identifier.urihttp://hdl.handle.net/10400.18/9050
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherOxford University Press/ UK Environmental Mutagen Societypt_PT
dc.relation.publisherversionhttps://academic.oup.com/mutage/article-abstract/38/5/273/7207507?redirectedFrom=fulltextpt_PT
dc.rights.urihttp://creativecommons.org/licenses/by-nc/4.0/pt_PT
dc.subjectComet Assaypt_PT
dc.subjectDNA Strand Breakspt_PT
dc.subjectCryopreservationpt_PT
dc.subjectGenotoxicitypt_PT
dc.subjectValidationpt_PT
dc.subjectGenotoxicidade Ambientalpt_PT
dc.titleDNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trialpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage282pt_PT
oaire.citation.issue5pt_PT
oaire.citation.startPage273pt_PT
oaire.citation.titleMutagenesispt_PT
oaire.citation.volume38pt_PT
rcaap.embargofctAcesso de acordo com política editorial da revista.pt_PT
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT

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