Tissue Sample Preparation for In Vivo Rodent Alkaline Comet Assay
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- Nakajima Madoka
- Biosafety Research Center, Foods, Drugs and Pesticides
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- Ueda Maya
- Biosafety Research Center, Foods, Drugs and Pesticides
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- Yamakage Kohji
- Hatano Research Institute, Food and Drug Safety Center
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- Nakagawa Yuzuki
- Hatano Research Institute, Food and Drug Safety Center
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- Nakagawa Munehiro
- Drug Development Service Segment, Mitsubishi Chemical Medience Co.
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- Ohyama Wakako
- Yakult Central Institute for Microbiological Research
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- Omori Takashi
- Doshisha University
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- Asano Norihide
- Kinki University
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- Hayashi Makoto
- Biosafety Research Center, Foods, Drugs and Pesticides
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- Uno Yoshifumi
- Safety Research Laboratories, Mitsubishi Tanabe Pharma Co.
Bibliographic Information
- Other Title
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- Tissue sample preparation for in vitro rodent alkaline Comet assay
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Abstract
The Japanese Environmental Mutagen Society/the Mammalian Mutagenicity Study group conducted a collaborative study to investigate whether cell nuclei or whole cells might be more suitably used to correctly detect genotoxic chemicals in the in vivo rodent alkaline Comet assay. Four participating laboratories applied four sample processing methods, i.e., three homogenization methods using the usual Potter-type shaft, a customized (loose) Potter-type shaft, or a Downs-loose-type shaft, for preparing cell nuclei, and the mesh membrane method for preparing whole cells, to the male rat liver. Homogenization with the usual Potter-type shaft clearly produced damage of the cell nuclei and DNA, while the other three methods seemed to provide similar conditions of the tissue samples. The proportion of cell nuclei: whole cells was 80-90%: 10-20% in all laboratories when the samples were prepared by homogenization using a Downs-loose-type shaft or by the mesh membrane method. The %DNA in tail were comparable in both samples among the negative control groups (single oral administration with physiological saline) of all laboratories, and showed an equal degree of increase in both samples of the ethyl methanesulfonate groups (single oral administration at 250 mg/kg) in all laboratories. In conclusion, the homogenization method using a loosely customized Potter-type shaft or a Downs-loose-type shaft, and the mesh membrane method would be equally acceptable for the in vivo rodent alkaline Comet assay.<br>
Journal
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- Genes and Environment
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Genes and Environment 34 (1), 50-54, 2012
The Japanese Environmental Mutagen Society
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Details 詳細情報について
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- CRID
- 1390001205257117184
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- NII Article ID
- 130004481196
- 10030121908
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- NII Book ID
- AA1212552X
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- COI
- 1:CAS:528:DC%2BC38XmtlGksLg%3D
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- ISSN
- 18807062
- 18807046
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- NDL BIB ID
- 023449025
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- CiNii Articles
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- Abstract License Flag
- Disallowed