DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trial
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , , , , , , , , , , , , , , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.18/9050 |
Resumo: | The 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. |
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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 trialComet AssayDNA Strand BreaksCryopreservationGenotoxicityValidationGenotoxicidade AmbientalThe 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.Certain 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.).Oxford University Press/ UK Environmental Mutagen SocietyRepositório Científico do Instituto Nacional de SaúdeMøller, PeterAzqueta, AmayaRodriguez-Garraus, AdrianaBakuradze, TamaraRichling, ElkeBankoglu, Ezgi EyluelStopper, HelgaClaudino Bastos, VictoriaLangie, Sabine A.S.Jensen, AnnieRistori, SaraScavone, FrancescaGiovannelli, LisaWojewódzka, MariaKruszewski, MarcinValdiglesias, VanessaLaffon, BlancaCosta, CarlaCosta, SolangePaulo Teixeira, JoãoMarino, MirkoDel Bo, CristianRiso, PatriziaZheng, CongyingShaposhnikov, SergeyCollins, Andrew2024-02-08T10:39:53Z2023-10-142023-10-14T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.18/9050engMutagenesis. 2023 Oct 14;38(5):273-282. doi: 10.1093/mutage/gead0190267-835710.1093/mutage/gead019metadata only accessinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-02-10T01:30:46Zoai:repositorio.insa.pt:10400.18/9050Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T02:37:21.661377Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trial |
title |
DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trial |
spellingShingle |
DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trial Møller, Peter Comet Assay DNA Strand Breaks Cryopreservation Genotoxicity Validation Genotoxicidade Ambiental |
title_short |
DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trial |
title_full |
DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trial |
title_fullStr |
DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trial |
title_full_unstemmed |
DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trial |
title_sort |
DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trial |
author |
Møller, Peter |
author_facet |
Møller, Peter Azqueta, Amaya Rodriguez-Garraus, Adriana Bakuradze, Tamara Richling, Elke Bankoglu, Ezgi Eyluel Stopper, Helga Claudino Bastos, Victoria Langie, Sabine A.S. Jensen, Annie Ristori, Sara Scavone, Francesca Giovannelli, Lisa Wojewódzka, Maria Kruszewski, Marcin Valdiglesias, Vanessa Laffon, Blanca Costa, Carla Costa, Solange Paulo Teixeira, João Marino, Mirko Del Bo, Cristian Riso, Patrizia Zheng, Congying Shaposhnikov, Sergey Collins, Andrew |
author_role |
author |
author2 |
Azqueta, Amaya Rodriguez-Garraus, Adriana Bakuradze, Tamara Richling, Elke Bankoglu, Ezgi Eyluel Stopper, Helga Claudino Bastos, Victoria Langie, Sabine A.S. Jensen, Annie Ristori, Sara Scavone, Francesca Giovannelli, Lisa Wojewódzka, Maria Kruszewski, Marcin Valdiglesias, Vanessa Laffon, Blanca Costa, Carla Costa, Solange Paulo Teixeira, João Marino, Mirko Del Bo, Cristian Riso, Patrizia Zheng, Congying Shaposhnikov, Sergey Collins, Andrew |
author2_role |
author author author author author author author author author author author author author author author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Repositório Científico do Instituto Nacional de Saúde |
dc.contributor.author.fl_str_mv |
Møller, Peter Azqueta, Amaya Rodriguez-Garraus, Adriana Bakuradze, Tamara Richling, Elke Bankoglu, Ezgi Eyluel Stopper, Helga Claudino Bastos, Victoria Langie, Sabine A.S. Jensen, Annie Ristori, Sara Scavone, Francesca Giovannelli, Lisa Wojewódzka, Maria Kruszewski, Marcin Valdiglesias, Vanessa Laffon, Blanca Costa, Carla Costa, Solange Paulo Teixeira, João Marino, Mirko Del Bo, Cristian Riso, Patrizia Zheng, Congying Shaposhnikov, Sergey Collins, Andrew |
dc.subject.por.fl_str_mv |
Comet Assay DNA Strand Breaks Cryopreservation Genotoxicity Validation Genotoxicidade Ambiental |
topic |
Comet Assay DNA Strand Breaks Cryopreservation Genotoxicity Validation Genotoxicidade Ambiental |
description |
The 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. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-10-14 2023-10-14T00:00:00Z 2024-02-08T10:39:53Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.18/9050 |
url |
http://hdl.handle.net/10400.18/9050 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Mutagenesis. 2023 Oct 14;38(5):273-282. doi: 10.1093/mutage/gead019 0267-8357 10.1093/mutage/gead019 |
dc.rights.driver.fl_str_mv |
metadata only access info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
metadata only access |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Oxford University Press/ UK Environmental Mutagen Society |
publisher.none.fl_str_mv |
Oxford University Press/ UK Environmental Mutagen Society |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
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1799137424790519808 |