Assessment of technical protocols for novel embryonic stem cell tests with molecular markers (Hand1- and Cmya1-ESTs)

• Published in: Journal of toxicological sciences Vol. 37; no. 4; p. 845
• Main Authors: Suzuki, Noriyuki, Yamashita, Norihisa, Koseki, Naoteru, Yamada, Toru, Kimura, Yutaka, Aiba, Setsuya, Toyoizumi, Tomoyasu, Watanabe, Mika, Ohta, Ryo, Tanaka, Noriho, Saito, Koichi
• Format: Journal Article
• Language: English
• Published: Suita Japan Science and Technology Agency 01.07.2012
• ISSN: 0388-1350; 1880-3989

The Hand1- and Cmya1-ESTs are novel short-term tests for embryotoxic chemicals using genetically engineering mouse ES cells for luciferase reporter gene assays. These ESTs allow convenient determination of differentiation toxicity and cell viability in a short duration with high throughput 96-well microplates for prediction of embryotoxicity of chemicals. To assess the Hand1-EST technical protocol, we firstly compared reporter gene assay and cytotoxicity test data for a representative compound (hydroxyurea) from four different laboratories with tests carried out under the same experimental conditions. Extensive investigations of the Hand1- and Cmya1-ESTs were then performed to explore reproducibility by comparing a set of 6 well-known test chemicals, including hydroxyurea, across the laboratories. The results gave good correspondence in all four laboratories, indicating that transferability, intra-laboratory variability and inter-laboratory variability of the present technical protocols of the ESTs were sufficient to conduct further validation studies.