Improvement of the embryonic stem cell test endpoint analysis by use of field potential detection
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概要
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The embryonic stem cell test (EST) is a validated in vitro method to assess the embryotoxic potential of compounds and is a promising tool for drug screening. EST requires microscopic observation of beating cardiomyocytes differentiated from embryonic stem cells as a toxicological endpoint. However, this process is time-consuming and lacks throughput performance. To improve the analysis, we introduced an electrophysiological method with a microelectrode array system for the evaluation of differentiated cardiomyocytes. Embryotoxic (valproic acid, verapamil, and 5-fluorouracil) and non-embryotoxic (penicillin G, d-camphor, and isoniazid) compounds were assessed with the system. Mouse embryonic stem cells were differentiated into cardiomyocytes and treated with each compound during the differentiation process. The embryotoxicity of each compound was then assessed by measuring the field potentials of differentiated cardiomyocytes using the microelectrode array system, as well as by microscopic evaluation. All the embryotoxic compounds dose-dependently inhibited the field potential formation and the myocardial beating of differentiated cells, while the non-embryotoxic compounds did not affect either endpoint. The detection capabilities of the two assay methods were similar. These results indicated that the field potential measurements can be used as an alternative endpoint of EST. Moreover, the field potential can be measured automatically, introducing a high throughput performance compared to the conventional microscopic observation. We therefore concluded that the endpoint analysis with the microelectrode array system improves the original EST and can be useful for the assessment of the embryotoxic potential of compounds.
- 2010-10-01
著者
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Funabashi Hitoshi
Safety Research Laboratories, Dainippon Sumitomo Pharma. Co., Ltd.
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Seki Takaki
Safety Research Laboratories, Dainippon Sumitomo Pharma. Co., Ltd.
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Seki Takaki
Safety Res. Laboratories Dainippon Sumitomo Pharma Co. Ltd.
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Sukata Tokuo
Environmental Health Science Lab. Sumitomo Chemical Co. Ltd.
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KOSEKI Naoteru
Safety Research Laboratories, Dainippon Sumitomo Pharma Co., Ltd.
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DEGUCHI Jiro
Safety Research Laboratories, Dainippon Sumitomo Pharma Co., Ltd.
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YAMADA Toru
Safety Research Laboratories, Dainippon Sumitomo Pharma Co., Ltd.
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Funabashi Hitoshi
Safety Research Laboratories Dainippon Sumitomo Pharma Co. Ltd.
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Yamada Toru
Safety Research Laboratories Dainippon Sumitomo Pharma Co. Ltd.
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Deguchi Jiro
Safety Research Laboratories Dainippon Sumitomo Pharma Co. Ltd.
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Koseki Naoteru
Safety Research Laboratories Dainippon Sumitomo Pharma Co. Ltd.
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Sukata Tokuo
Safety Research Laboratories, Dainippon Sumitomo Pharma Co., Ltd.
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