Inter-laboratory Validation of the in vivo Flow Cytometric icronucleus Analysis Method (MicroFlow(R)

• 作 者: Yan Chang Zhenna Xia Zhaomin Zhong Ming Zhang Fengjun An Yiyi Cao Xingchao Geng Ying Jiang Qing Ju Yongsheng Yu Jiangbo Zhu Changhui Zhou Stephen D.Dertinger Bo Li Mingyang Liao Bojun Yuan Tianbao Zha
• 机 构: National Shanghai Center for New Drug Safety Evaluation & Research.China State Institute of Pharmace
• 出 处: 中国环境诱变剂学会致突变、致畸学术讨论会
• 发表日期: 2014年5期

　　Although inter-laboratory validation efforts of the flow cytometry (FCM) based in vivo micronucleus (MN) assay have taken place in the EU and US, none have been organized in China.Therefore, an inter-laboratory study that included eight laboratories in China and one experienced reference laboratory in the US was coordinated to validate the in vivo FCM MicroFlow(R) method for enumerating micronucleated reticulocytes (MN-RETs) in rat blood.Assay reliability and reproducibility were evaluated using four known genotoxicants, and results obtained using the FCM method were compared with results from the traditional bone marrow evaluation using microscopy.Each of the four chemicals was tested at 3 sites (2 in China and the one US reference laboratory).After three consecutive daily exposures to a genotoxicant, blood and bone marrow samples were obtained from rats 24 hours after the third dose.MN-RET frequencies were measured in 20,000 RET in blood by FCM and micronucleated polychromatic erythrocyte (MN-PCE) frequencies were measured in 2,000 PCEs in bone marrow by microscopy.For both methods of analysis, each genotoxicant was shown to induce a statistically significant increase in the frequency of MN in at least one dose.Where more than one doses caused increases, responsesoccurred in a dose-dependent manner.Spearmanscorrelation coefficient (rs) for FCM-based MN-RET versus microscopy-based MN-PCE measurements (eightexperiments, 200 paired measurements) was0.723, indicating a high degree of correspondencebetween methods and compartments.The rs value for replicate flowcytometric MN-RET measurements performed at the eightcollaborative laboratories was 0.940 (n =200), and between the 8 FCM laboratories with the reference laboratory was 0.933 (n =200), suggesting that the automated method is highlytransferable between laboratories.The flow cytometric micronucleus analysis method is currently used in many countries worldwide, and these data support its use for evaluating the in vivo genotoxic potential of test chemicals in China.