Assessment of technical protocols for novel embryonic stem cell tests with molecular markers (Hand1- and Cmya1-ESTs): A preliminary cross-laboratory performance analysis

Noriyuki Suzuki, Norihisa Yamashita, Naoteru Koseki, Toru Yamada, Yutaka Kimura, Setsuya Aiba, Tomoyasu Toyoizumi, Mika Watanabe, Ryo Ohta, Noriho Tanaka, Koichi Saito

Research output: Contribution to journalLetter

13 Citations (Scopus)

Abstract

The Hand1- and Cmya1-ESTs are novel short-term tests for embryotoxic chemicalsusing 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.

Original languageEnglish
Pages (from-to)845-851
Number of pages7
JournalJournal of Toxicological Sciences
Volume37
Issue number4
DOIs
Publication statusPublished - 2012

Keywords

  • Differentiation
  • Embryonic stem cell test
  • Embryonic stem cells
  • Embryotoxicity
  • In vitro
  • Reporter gene assay

ASJC Scopus subject areas

  • Toxicology

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