Application of dicentric chromosome assay for evaluation of radioprotective effect
The dicentric chromosome assay is a well-established biodosimetric method used to assess absorbed ionizing radiation doses by detecting dicentric chromosomal aberrations. Here, we present a detailed, reproducible protocol for applying the dicentric chromosome assay for in vitro evaluation of radiopr...
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| Published in | Biology methods and protocols Vol. 10; no. 1; p. bpaf058 |
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| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
| Published |
England
Oxford University Press
2025
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| Subjects | |
| Online Access | Get full text |
| ISSN | 2396-8923 2396-8923 |
| DOI | 10.1093/biomethods/bpaf058 |
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| Abstract | The dicentric chromosome assay is a well-established biodosimetric method used to assess absorbed ionizing radiation doses by detecting dicentric chromosomal aberrations. Here, we present a detailed, reproducible protocol for applying the dicentric chromosome assay for in vitro evaluation of radioprotective agents, including novel piperazine derivatives compared with amifostine and its active metabolite WR-1065. The protocol covers all key steps—blood sample preparation, in vitro irradiation, lymphocyte culture, metaphase preparation, and scoring of dicentric chromosomes. It highlights critical stages that affect data quality and reproducibility. Integrating manual scoring with automated analysis using the Metafer system ensures accurate and efficient assessment. Thus, this protocol bridges the fields of biological dosimetry and preclinical screening of radioprotective agents, providing a reliable framework for emergency radiation dose estimation and the development of new radiation medical countermeasures. |
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| AbstractList | The dicentric chromosome assay is a well-established biodosimetric method used to assess absorbed ionizing radiation doses by detecting dicentric chromosomal aberrations. Here, we present a detailed, reproducible protocol for applying the dicentric chromosome assay for
evaluation of radioprotective agents, including novel piperazine derivatives compared with amifostine and its active metabolite WR-1065. The protocol covers all key steps-blood sample preparation,
irradiation, lymphocyte culture, metaphase preparation, and scoring of dicentric chromosomes. It highlights critical stages that affect data quality and reproducibility. Integrating manual scoring with automated analysis using the Metafer system ensures accurate and efficient assessment. Thus, this protocol bridges the fields of biological dosimetry and preclinical screening of radioprotective agents, providing a reliable framework for emergency radiation dose estimation and the development of new radiation medical countermeasures. The dicentric chromosome assay is a well-established biodosimetric method used to assess absorbed ionizing radiation doses by detecting dicentric chromosomal aberrations. Here, we present a detailed, reproducible protocol for applying the dicentric chromosome assay for in vitro evaluation of radioprotective agents, including novel piperazine derivatives compared with amifostine and its active metabolite WR-1065. The protocol covers all key steps—blood sample preparation, in vitro irradiation, lymphocyte culture, metaphase preparation, and scoring of dicentric chromosomes. It highlights critical stages that affect data quality and reproducibility. Integrating manual scoring with automated analysis using the Metafer system ensures accurate and efficient assessment. Thus, this protocol bridges the fields of biological dosimetry and preclinical screening of radioprotective agents, providing a reliable framework for emergency radiation dose estimation and the development of new radiation medical countermeasures. The dicentric chromosome assay is a well-established biodosimetric method used to assess absorbed ionizing radiation doses by detecting dicentric chromosomal aberrations. Here, we present a detailed, reproducible protocol for applying the dicentric chromosome assay for in vitro evaluation of radioprotective agents, including novel piperazine derivatives compared with amifostine and its active metabolite WR-1065. The protocol covers all key steps—blood sample preparation, in vitro irradiation, lymphocyte culture, metaphase preparation, and scoring of dicentric chromosomes. It highlights critical stages that affect data quality and reproducibility. Integrating manual scoring with automated analysis using the Metafer system ensures accurate and efficient assessment. Thus, this protocol bridges the fields of biological dosimetry and preclinical screening of radioprotective agents, providing a reliable framework for emergency radiation dose estimation and the development of new radiation medical countermeasures. The dicentric chromosome assay is a well-established biodosimetric method used to assess absorbed ionizing radiation doses by detecting dicentric chromosomal aberrations. Here, we present a detailed, reproducible protocol for applying the dicentric chromosome assay for in vitro evaluation of radioprotective agents, including novel piperazine derivatives compared with amifostine and its active metabolite WR-1065. The protocol covers all key steps-blood sample preparation, in vitro irradiation, lymphocyte culture, metaphase preparation, and scoring of dicentric chromosomes. It highlights critical stages that affect data quality and reproducibility. Integrating manual scoring with automated analysis using the Metafer system ensures accurate and efficient assessment. Thus, this protocol bridges the fields of biological dosimetry and preclinical screening of radioprotective agents, providing a reliable framework for emergency radiation dose estimation and the development of new radiation medical countermeasures.The dicentric chromosome assay is a well-established biodosimetric method used to assess absorbed ionizing radiation doses by detecting dicentric chromosomal aberrations. Here, we present a detailed, reproducible protocol for applying the dicentric chromosome assay for in vitro evaluation of radioprotective agents, including novel piperazine derivatives compared with amifostine and its active metabolite WR-1065. The protocol covers all key steps-blood sample preparation, in vitro irradiation, lymphocyte culture, metaphase preparation, and scoring of dicentric chromosomes. It highlights critical stages that affect data quality and reproducibility. Integrating manual scoring with automated analysis using the Metafer system ensures accurate and efficient assessment. Thus, this protocol bridges the fields of biological dosimetry and preclinical screening of radioprotective agents, providing a reliable framework for emergency radiation dose estimation and the development of new radiation medical countermeasures. |
| Author | Chmil, Vojtěch Lecová, Lenka Tichý, Aleš Milanová, Marcela |
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| Cites_doi | 10.14309/ctg.0000000000000347 10.1039/D4MD00311J 10.1007/s11894-011-0214-8 10.51083/radiatenvironmed.11.2_91 10.1667/RADE-22-00202.1 10.3109/09553002.2016.1144941 10.1038/s41572-019-0064-5 10.1016/S0140-6736(00)02176-0 10.1016/j.jmir.2015.10.006 10.1667/RADE-22-00207.1 10.1159/000228561 10.1080/09553002.2023.2173822 |
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| Copyright | The Author(s) 2025. Published by Oxford University Press. The Author(s) 2025. Published by Oxford University Press. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. The Author(s) 2025. Published by Oxford University Press. 2025 |
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| Keywords | chromosome aberrations amifostine radioprotective agent cytogenetics ionizing radiation |
| Language | English |
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| References | Endesfelder (2025081318460764000_bpaf058-B10) 2023; 199 Agency (2025081318460764000_bpaf058-B8) 2011 Chmil (2025081318460764000_bpaf058-B7) 2024; 15 Friedman (2025081318460764000_bpaf058-B13) 2016; 92 Chang (2025081318460764000_bpaf058-B2) 2011; 13 Nakayama (2025081318460764000_bpaf058-B9) 2022; 11 Port (2025081318460764000_bpaf058-B11) 2023; 199 Chmil (2025081318460764000_bpaf058-B6) 2023; 99 Holdstock (2025081318460764000_bpaf058-B5) 2000; 355 Thiruvengadam (2025081318460764000_bpaf058-B1) 2021; 12 Herfarth (2025081318460764000_bpaf058-B3) 2009; 27 Pirayesh Islamian (2025081318460764000_bpaf058-B12) 2016; 47 De Ruysscher (2025081318460764000_bpaf058-B4) 2019; 5 |
| References_xml | – volume: 12 start-page: e00347 year: 2021 ident: 2025081318460764000_bpaf058-B1 article-title: Burden of ionizing radiation in the diagnosis and management of necrotizing pancreatitis publication-title: Clin Transl Gastroenterol doi: 10.14309/ctg.0000000000000347 – volume: 15 start-page: 2855 year: 2024 ident: 2025081318460764000_bpaf058-B7 article-title: Second-generation piperazine derivatives as promising radiation countermeasures publication-title: RSC Med Chem doi: 10.1039/D4MD00311J – volume: 13 start-page: 449 year: 2011 ident: 2025081318460764000_bpaf058-B2 article-title: Cancer risk related to gastrointestinal diagnostic radiation exposure publication-title: Curr Gastroenterol Rep doi: 10.1007/s11894-011-0214-8 – volume: 11 start-page: 91 year: 2022 ident: 2025081318460764000_bpaf058-B9 article-title: Cytogenetic biodosimetry in radiation emergency medicine: 4. Overview of cytogenetic biodosimetry publication-title: Radiat Environ Med doi: 10.51083/radiatenvironmed.11.2_91 – volume: 199 start-page: 556 year: 2023 ident: 2025081318460764000_bpaf058-B10 article-title: RENEB Inter-Laboratory Comparison 2021: the dicentric chromosome assay publication-title: Radiat Res doi: 10.1667/RADE-22-00202.1 – volume: 92 start-page: 229 year: 2016 ident: 2025081318460764000_bpaf058-B13 article-title: Influence of nuclear structure on the formation of radiation-induced lethal lesions publication-title: Int J Radiat Biol doi: 10.3109/09553002.2016.1144941 – volume: 5 start-page: 13 year: 2019 ident: 2025081318460764000_bpaf058-B4 article-title: Radiotherapy toxicity publication-title: Nat Rev Dis Primers doi: 10.1038/s41572-019-0064-5 – volume: 355 start-page: 1544 year: 2000 ident: 2025081318460764000_bpaf058-B5 article-title: Nuclear weapons, a continuing threat to health publication-title: Lancet doi: 10.1016/S0140-6736(00)02176-0 – volume: 47 start-page: 49 year: 2016 ident: 2025081318460764000_bpaf058-B12 article-title: Radioprotective effects of amifostine and lycopene on human peripheral blood lymphocytes in vitro publication-title: J Med Imaging Radiat Sci doi: 10.1016/j.jmir.2015.10.006 – volume: 199 start-page: 535 year: 2023 ident: 2025081318460764000_bpaf058-B11 article-title: RENEB Inter-Laboratory Comparison 2021: inter-assay comparison of eight dosimetry assays publication-title: Radiat Res doi: 10.1667/RADE-22-00207.1 – start-page: 1 volume-title: Cytogenetic Dosimetry: Applications in Preparedness for and Response to Radiation Emergencies year: 2011 ident: 2025081318460764000_bpaf058-B8 – volume: 27 start-page: 278 year: 2009 ident: 2025081318460764000_bpaf058-B3 article-title: Risk of radiation and choice of imaging publication-title: Dig Dis doi: 10.1159/000228561 – volume: 99 start-page: 1148 year: 2023 ident: 2025081318460764000_bpaf058-B6 article-title: Looking for the Phoenix: the current research on radiation countermeasures publication-title: Int J Radiat Biol doi: 10.1080/09553002.2023.2173822 |
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| SubjectTerms | Amifostine Chromosome aberrations Chromosomes Dosimetry Ionizing radiation Lymphocytes Metaphase Nuclear accidents & safety Piperazine Protocols Radioprotective agents Software |
| Title | Application of dicentric chromosome assay for evaluation of radioprotective effect |
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