Farabaugh Christopher S, Doak Shareen, Roy Shambhu, Elespuru Rosalie
Charles River Laboratories, Skokie, IL, United States.
Institute of Life Science 2, Swansea University Medical School, Singleton Park, Swansea, Wales, United Kingdom.
Front Toxicol. 2023 Apr 26;5:1171960. doi: 10.3389/ftox.2023.1171960. eCollection 2023.
The micronucleus (MNvit) assay is used to evaluate the aneugenic and clastogenic potential of a test material based upon its ability to induce micronuclei in the cells. This protocol is provided for testing of nanomaterials (NM) with standard cell lines in the absence of metabolic activation. The use of cytochalasin B (CytoB) and the analysis of binucleated cells in the cytokinesis-block version of the micronucleus assay ensures that cells analyzed have undergone cell division, which is required for expression of DNA damage and micronucleus formation. Issues specific to NM that were problematic with standard test methods are addressed, including test system choice, dose selection, test material exposures, CytoB timing, cytotoxicity determination, and DNA damage expression time. A step-by-step protocol for micronucleus assessment of NM is provided.
微核（MNvit）试验用于根据测试材料诱导细胞微核的能力来评估其产生非整倍体和断裂剂的潜力。本方案用于在无代谢活化情况下用标准细胞系测试纳米材料（NM）。在微核试验的胞质分裂阻滞版本中使用细胞松弛素B（CytoB）并分析双核细胞，可确保所分析的细胞已经历细胞分裂，这是DNA损伤表达和微核形成所必需的。解决了标准测试方法中存在问题的NM特有的问题，包括测试系统选择、剂量选择、测试材料暴露、CytoB时机、细胞毒性测定和DNA损伤表达时间。提供了NM微核评估的分步方案。
