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Effect of cryopreservation on DNA damage and DNA repair activity in human blood samples in the comet assay
Zitieren Sie bitte immer diese URN: urn:nbn:de:bvb:20-opus-265326
- The comet assay is a commonly used method to determine DNA damage and repair activity in many types of samples. In recent years, the use of the comet assay in human biomonitoring became highly attractive due to its various modified versions, which may be useful to determine individual susceptibility in blood samples. However, in human biomonitoring studies, working with large sample numbers that are acquired over an extended time period requires some additional considerations. One of the most important issues is the storage of samples and itsThe comet assay is a commonly used method to determine DNA damage and repair activity in many types of samples. In recent years, the use of the comet assay in human biomonitoring became highly attractive due to its various modified versions, which may be useful to determine individual susceptibility in blood samples. However, in human biomonitoring studies, working with large sample numbers that are acquired over an extended time period requires some additional considerations. One of the most important issues is the storage of samples and its effect on the outcome of the comet assay. Another important question is the suitability of different blood preparations. In this study, we analysed the effect of cryopreservation on DNA damage and repair activity in human blood samples. In addition, we investigated the suitability of different blood preparations. The alkaline and FPG as well as two different types of repair comet assay and an in vitro hydrogen peroxide challenge were applied. Our results confirmed that cryopreserved blood preparations are suitable for investigating DNA damage in the alkaline and FPG comet assay in whole blood, buffy coat and PBMCs. Ex vivo hydrogen peroxide challenge yielded its optimal effect in isolated PBMCs. The utilised repair comet assay with either UVC or hydrogen peroxide-induced lesions and an aphidicolin block worked well in fresh PBMCs. Cryopreserved PBMCs could not be used immediately after thawing. However, a 16-h recovery with or without mitotic stimulation enabled the application of the repair comet assay, albeit only in a surviving cell fraction.…
Autor(en): | Ezgi Eyluel Bankoglu, Franzisca Stipp, Johanna Gerber, Florian Seyfried, August Heidland, Udo Bahner, Helga Stopper |
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URN: | urn:nbn:de:bvb:20-opus-265326 |
Dokumentart: | Artikel / Aufsatz in einer Zeitschrift |
Institute der Universität: | Medizinische Fakultät / Klinik und Poliklinik für Allgemein-, Viszeral-, Gefäß- und Kinderchirurgie (Chirurgische Klinik I) |
Medizinische Fakultät / Institut für Pharmakologie und Toxikologie | |
Sprache der Veröffentlichung: | Englisch |
Titel des übergeordneten Werkes / der Zeitschrift (Englisch): | Archives of Toxicology |
Erscheinungsjahr: | 2021 |
Band / Jahrgang: | 95 |
Heft / Ausgabe: | 5 |
Seitenangabe: | 1831–1841 |
Originalveröffentlichung / Quelle: | Archives of Toxicology 2021, 95(5):1831–1841. DOI: 10.1007/s00204-021-03012-4 |
DOI: | https://doi.org/10.1007/s00204-021-03012-4 |
Allgemeine fachliche Zuordnung (DDC-Klassifikation): | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
Freie Schlagwort(e): | DNA damage; DNA repair; blood samples; comet assay; human biomonitoring |
Datum der Freischaltung: | 12.04.2022 |
Lizenz (Deutsch): | ![]() |