Comparative study of three cell morphological analysis methods on in vitro micronucleus assay
10.19428/j.cnki.sjpm.2019.19680
- VernacularTitle:三种细胞形态学分析方法在体外微核检测中的比较研究
- Author:
Jing-qiu SUN
1
;
Yi SHUAI
;
Ping XIAO
;
Wei-dong ZHENG
;
Yan-qin WANG
;
Gong-hua TAO
Author Information
1. Shanghai Municipal Center for Disease Control and Prevention, Shanghai 200336, China
- Publication Type:Research Article
- Keywords:
in vitro micronucleus;
high-throughput screening;
flow cytometry;
high-content screening
- From:
Shanghai Journal of Preventive Medicine
2020;32(1):78-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the feasibility of genotoxicity assessment for chemicals via flow cytometry (FCM) and high-content screening (HCS) based on high-throughput screening in vitro micronucleus assays. Methods In reference to the methodology of OECD TG487, the typical positive controls, cyclophosphamide (CP) and mitomycin C (MMC), were selected.And no serum MEM medium was treated as negative control.Dose range of CP was 5-20 mg/L and MMC was 0.25-1.0 mg/L.CHL cells were treated with three concentrations of each chemical for 4 h.High-throughput screening in vitro micronucleus assays based on FCM and HCS were established.The results of the frequency of micronuclei were compared to traditional cytokinesis blocking micronucleus assay in each group with or without metabolic activation. Results The frequencies of micronuclei induced by CP and MMC (ascending rank) were separately 1.9%, 7.6%, 10.4% and 5.9%, 11.4%, 16.7%, which were obtained by conventional microscopic scoring.The frequencies of micronuclei induced by CP and MMC (ascending rank) were separately 2.8%, 2.6%, 7.8% and 3.2%, 3.7%, 5.1%, which were obtained by flow cytometry screening.The frequencies of micronuclei induced by CP and MMC (ascending rank) were separately2.8%, 6.2%, 9.1% and 7.9%, 10.1%, 10.2%, which were obtained by high-content screening.Compared with negative controls, the differences of the results were statistically significant(P < 0.05), and there was a dose-response relationship. Conclusion In this study, the results of high-throughput screening assays of FCM and HCS are in accordance to the results of traditional cytokinesis blocking micronucleus assay, indicating that high-throughput screening in vitro micronucleus assays could detect micronucleus formation automatically and improve the efficiency.Therefore, the method could provide data support for using high-throughput screening in vitro micronucleus assays into genotoxicity assessment of chemicals.
