title: Gene-modified embryonic stem cell test to characterize chemical risks.
abstract: A high-throughput test of cell growth inhibition was performed using mouse embryonic stem (ES) cells to assess chemical toxicities. We herein demonstrated using a 96-well culture plate approach and the MTT assay that this method was suitable for prioritization of chemicals for their cytotoxic properties. In order to categorize chemicals, we used p53 gene-modified mouse ES cells as well as wild-type ES cells. The p53 gene is a well-known tumor suppressor and controls programmed cell death (apoptosis) and cellular senescence that is triggered by DNA-damaging agents such as alkylating agents and radiation. In the present study, p53-deficient ES cells were found to be more resistant to a tumor initiator, diethylnitrosamine (DEN), than wild-type ES cells, suggesting the inhibition of apoptosis or senescence by a dysfunction in p53. Chromosome aberrations were more frequently detected in p53-deficient ES cells than in wild-type cells, indicating genomic instability due to the deletion of p53. Other tumor initiators, methyl methanesulfonate (MMS) and N-methyl-N-nitrosourea (NMU), did not reveal apparent differences in cytotoxicity between wild-type and p53-deficient ES cells. Thus, ES test system using gene-modified ES cells may be used to categorize chemicals by detecting their characteristic effects on apoptosis, genotoxic potentials as well as general cytotoxicity.
authors.affiliation: Department of Obstetrics and Gynecology, Osaka City University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka, 545-8585, Japan.
authors.forename: Kohei
authors.initials: K
authors.lastname: Kitada
authors.affiliation: Department of Molecular Genetics, Osaka City University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka, 545-8585, Japan.
authors.forename: Akane
authors.initials: A
authors.lastname: Kizu
authors.affiliation: Institute of Advanced Clinical Medicine, Faculty of Medicine, Kindai University, 377-2 Onohigashi, Sayama, Osaka, 589-8511, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Teramura
authors.affiliation: Institute of Advanced Clinical Medicine, Faculty of Medicine, Kindai University, 377-2 Onohigashi, Sayama, Osaka, 589-8511, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Takehara
authors.affiliation: Department of Obstetrics and Gynecology, Osaka City University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka, 545-8585, Japan.
authors.forename: Masami
authors.initials: M
authors.lastname: Hayashi
authors.affiliation: Department of Obstetrics and Gynecology, Osaka City University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka, 545-8585, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Tachibana
authors.affiliation: Department of Pathology, Osaka City University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka, 545-8585, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Wanibuchi
authors.affiliation: Japan Bioassay Research Center, 2445 Hirasawa, Hadano, Kanagawa, 257-0015, Japan.
authors.forename: Shoji
authors.initials: S
authors.lastname: Fukushima
authors.affiliation: Department of Obstetrics and Gynecology, Osaka City University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka, 545-8585, Japan.
authors.forename: Masayasu
authors.initials: M
authors.lastname: Koyama
authors.affiliation: Department of Molecular Genetics, Osaka City University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka, 545-8585, Japan. k-yoshida@med.osaka-cu.ac.jp.
authors.forename: Kayo
authors.initials: K
authors.lastname: Yoshida
authors.affiliation: Department of Molecular Genetics, Osaka City University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka, 545-8585, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Morita
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics*; D002478:Cells, Cultured; D004249:DNA Damage; D004052:Diethylnitrosamine / Q000633:toxicity; D053595:Embryonic Stem Cells / Q000187:drug effects*; D051379:Mice; D018675:Toxicity Tests / Q000379:methods*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
chemical_list: D016159:Tumor Suppressor Protein p53; D004052:Diethylnitrosamine
keywords: Chemical risks; Cytotoxicity; Gene modification; High-throughput analysis; Mouse embryonic stem cells; p53