Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies
Autor(a) principal: | |
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Data de Publicação: | 2021 |
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/10400.21/13625 |
Resumo: | DNA damage and repair activity are often assessed in blood samples from humans in different types of molecular epidemiology studies. However, it is not always feasible to analyze the samples on the day of collection without any type of storage. For instance, certain studies use repeated sampling of cells from the same subject or samples from different subjects collected at different time points, and it is desirable to analyze all these samples in the same comet assay experiment. In addition, flawless comet assay analyses on frozen samples open up the possibility of using this technique on biobank material. In this article, we discuss the use of cryopreserved peripheral blood mononuclear cells (PBMCs), buffy coat (BC), and whole blood (WB) for analysis of DNA damage and repair using the comet assay. The published literature and the authors' experiences indicate that various types of blood samples can be cryopreserved with only a minor effect on the basal level of DNA damage. There is evidence to suggest that WB and PBMCs can be cryopreserved for several years without much effect on the level of DNA damage. However, care should be taken when cryopreserving WB and BCs. It is possible to use either fresh or frozen samples of blood cells, but results from fresh and frozen cells should not be used in the same dataset. The article outlines detailed protocols for the cryopreservation of PBMCs, BCs, and WB samples. |
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Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studiesComet assayCryopreservationDNADNA damageEpidemiologyMolecularLeucocytesWhole bloodDNA damage and repair activity are often assessed in blood samples from humans in different types of molecular epidemiology studies. However, it is not always feasible to analyze the samples on the day of collection without any type of storage. For instance, certain studies use repeated sampling of cells from the same subject or samples from different subjects collected at different time points, and it is desirable to analyze all these samples in the same comet assay experiment. In addition, flawless comet assay analyses on frozen samples open up the possibility of using this technique on biobank material. In this article, we discuss the use of cryopreserved peripheral blood mononuclear cells (PBMCs), buffy coat (BC), and whole blood (WB) for analysis of DNA damage and repair using the comet assay. The published literature and the authors' experiences indicate that various types of blood samples can be cryopreserved with only a minor effect on the basal level of DNA damage. There is evidence to suggest that WB and PBMCs can be cryopreserved for several years without much effect on the level of DNA damage. However, care should be taken when cryopreserving WB and BCs. It is possible to use either fresh or frozen samples of blood cells, but results from fresh and frozen cells should not be used in the same dataset. The article outlines detailed protocols for the cryopreservation of PBMCs, BCs, and WB samples.Oxford AcademicRCIPLMøller, PeterBankoglu, Ezgi EyluelStopper, HelgaGiovannelli, LisaLadeira, CarinaKoppen, GudrunGajski, GoranCollins, AndrewValdiglesias, VanessaLaffon, BlancaBoutet-Robinet, ElisaPerdry, HervéDel Bo’, CristianLangie, Sabine A SDusinska, MariaAzqueta, Amaya2021-052021-05-01T00:00:00Z2024-08-05T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.21/13625engMøller P, Bankoglu EE, Stopper H, Giovannelli L, Ladeira C, Koppen G, et al. Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies. Mutagenesis. 2021;36(3):193-212.10.1093/mutage/geab012info:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-08-03T10:08:39Zoai:repositorio.ipl.pt:10400.21/13625Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:21:31.993081Repositório 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 |
Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies |
title |
Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies |
spellingShingle |
Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies Møller, Peter Comet assay Cryopreservation DNA DNA damage Epidemiology Molecular Leucocytes Whole blood |
title_short |
Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies |
title_full |
Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies |
title_fullStr |
Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies |
title_full_unstemmed |
Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies |
title_sort |
Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies |
author |
Møller, Peter |
author_facet |
Møller, Peter Bankoglu, Ezgi Eyluel Stopper, Helga Giovannelli, Lisa Ladeira, Carina Koppen, Gudrun Gajski, Goran Collins, Andrew Valdiglesias, Vanessa Laffon, Blanca Boutet-Robinet, Elisa Perdry, Hervé Del Bo’, Cristian Langie, Sabine A S Dusinska, Maria Azqueta, Amaya |
author_role |
author |
author2 |
Bankoglu, Ezgi Eyluel Stopper, Helga Giovannelli, Lisa Ladeira, Carina Koppen, Gudrun Gajski, Goran Collins, Andrew Valdiglesias, Vanessa Laffon, Blanca Boutet-Robinet, Elisa Perdry, Hervé Del Bo’, Cristian Langie, Sabine A S Dusinska, Maria Azqueta, Amaya |
author2_role |
author author author author author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
RCIPL |
dc.contributor.author.fl_str_mv |
Møller, Peter Bankoglu, Ezgi Eyluel Stopper, Helga Giovannelli, Lisa Ladeira, Carina Koppen, Gudrun Gajski, Goran Collins, Andrew Valdiglesias, Vanessa Laffon, Blanca Boutet-Robinet, Elisa Perdry, Hervé Del Bo’, Cristian Langie, Sabine A S Dusinska, Maria Azqueta, Amaya |
dc.subject.por.fl_str_mv |
Comet assay Cryopreservation DNA DNA damage Epidemiology Molecular Leucocytes Whole blood |
topic |
Comet assay Cryopreservation DNA DNA damage Epidemiology Molecular Leucocytes Whole blood |
description |
DNA damage and repair activity are often assessed in blood samples from humans in different types of molecular epidemiology studies. However, it is not always feasible to analyze the samples on the day of collection without any type of storage. For instance, certain studies use repeated sampling of cells from the same subject or samples from different subjects collected at different time points, and it is desirable to analyze all these samples in the same comet assay experiment. In addition, flawless comet assay analyses on frozen samples open up the possibility of using this technique on biobank material. In this article, we discuss the use of cryopreserved peripheral blood mononuclear cells (PBMCs), buffy coat (BC), and whole blood (WB) for analysis of DNA damage and repair using the comet assay. The published literature and the authors' experiences indicate that various types of blood samples can be cryopreserved with only a minor effect on the basal level of DNA damage. There is evidence to suggest that WB and PBMCs can be cryopreserved for several years without much effect on the level of DNA damage. However, care should be taken when cryopreserving WB and BCs. It is possible to use either fresh or frozen samples of blood cells, but results from fresh and frozen cells should not be used in the same dataset. The article outlines detailed protocols for the cryopreservation of PBMCs, BCs, and WB samples. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-05 2021-05-01T00:00:00Z 2024-08-05T00:00:00Z |
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/10400.21/13625 |
url |
http://hdl.handle.net/10400.21/13625 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Møller P, Bankoglu EE, Stopper H, Giovannelli L, Ladeira C, Koppen G, et al. Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies. Mutagenesis. 2021;36(3):193-212. 10.1093/mutage/geab012 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/embargoedAccess |
eu_rights_str_mv |
embargoedAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Oxford Academic |
publisher.none.fl_str_mv |
Oxford Academic |
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 |
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RCAAP |
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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 |
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