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http://dx.doi.org/10.5338/KJEA.2018.37.3.27

Study on Optimization of Cytoplasm Conditions for In Vitro Micronucleus Test Using Chinese Hamster Lung Cells  

Paik, Min Kyoung (Chemical Safety Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Kim, Areumnuri (Chemical Safety Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Shin, Hye Rim (Chemical Safety Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Chon, Kyongmi (Chemical Safety Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Park, Kyung-Hun (Chemical Safety Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Ryu, Ji-Hyeok (Chemical Safety Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Moon, Byeong Chul (Chemical Safety Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Publication Information
Korean Journal of Environmental Agriculture / v.37, no.3, 2018 , pp. 229-234 More about this Journal
Abstract
BACKGROUND: in vitro micronucleus test (vitMNT) is one of the promising alternative testing methods in genotoxicity test and was adopted as OECD test guideline for chemical registration. This study was conducted to optimize the cytoplasm conditions in vitMNT using Chinese hamster lung (CHL) cell. METHODS AND RESULTS: In this study cytokinesis-block micronucleus test was conducted. Mitomycin C and colchicine were used as positive control chemicals and were treated for three hours (short time) or twenty-four hours (long time). Giemsa solution was used for cell staining. For optimization of vitMNT, the final fixative was prepared as five concentrations (0%, 1%, 3%, 5%, and 25%) of acetic acid in methanol, and treatment times of the final fixative were varied under four conditions (immediately, one hour, four hours, and one day). CONCLUSION: Acetic acid at 1% in methanol as the final fixative was most adequate to preserve the cytoplasm around the nucleus in the interphase cells. Also, fixative treatment time of cell suspension for one to four hours may minimize the cell rupture. These results can be helpful for getting an accurate result promptly due to clear visual distinction to score micronucleus in vitMNT using giemsa solution.
Keywords
Cytoplasm conditions; In vitro micronucleus test; Optimization;
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