title: Assessment of technical protocols for novel embryonic stem cell tests with molecular markers (Hand1- and Cmya1-ESTs): a preliminary cross-laboratory performance analysis.
abstract: 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.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Suzuki
authors.forename: Norihisa
authors.initials: N
authors.lastname: Yamashita
authors.forename: Naoteru
authors.initials: N
authors.lastname: Koseki
authors.forename: Toru
authors.initials: T
authors.lastname: Yamada
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Kimura
authors.forename: Setsuya
authors.initials: S
authors.lastname: Aiba
authors.forename: Tomoyasu
authors.initials: T
authors.lastname: Toyoizumi
authors.forename: Mika
authors.initials: M
authors.lastname: Watanabe
authors.forename: Ryo
authors.initials: R
authors.lastname: Ohta
authors.forename: Noriho
authors.initials: N
authors.lastname: Tanaka
authors.forename: Koichi
authors.initials: K
authors.lastname: Saito
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics*; D002454:Cell Differentiation; D003598:Cytoskeletal Proteins; D004268:DNA-Binding Proteins / Q000235:genetics*; D053595:Embryonic Stem Cells / Q000166:cytology*; D023381:Endpoint Determination; D020224:Expressed Sequence Tags / Q000737:chemistry; D005819:Genetic Markers; D007753:Laboratories; D051379:Mice; D009687:Nuclear Proteins / Q000235:genetics*; D015203:Reproducibility of Results
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D003598:Cytoskeletal Proteins; D004268:DNA-Binding Proteins; D005819:Genetic Markers; D009687:Nuclear Proteins; C572128:CMYA1 protein, mouse; C095572:Hand1 protein, mouse