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Gene-modified embryonic stem cell test to characterize chemical risks

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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.

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Acknowledgment

This work was partly supported by the Long-range Research Initiative (LRI) program of Japan Chemical Industry Association.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Osaka City University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka, 545-8585, Japan

    Kohei Kitada, Masami Hayashi, Daisuke Tachibana & Masayasu Koyama

  2. Department of Molecular Genetics, Osaka City University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka, 545-8585, Japan

    Akane Kizu, Kayo Yoshida & Takashi Morita

  3. Institute of Advanced Clinical Medicine, Faculty of Medicine, Kindai University, 377-2 Onohigashi, Sayama, Osaka, 589-8511, Japan

    Takeshi Teramura & Toshiyuki Takehara

  4. Department of Pathology, Osaka City University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka, 545-8585, Japan

    Hideki Wanibuchi

  5. Japan Bioassay Research Center, 2445 Hirasawa, Hadano, Kanagawa, 257-0015, Japan

    Shoji Fukushima

Authors
  1. Kohei Kitada
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  2. Akane Kizu
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  3. Takeshi Teramura
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  4. Toshiyuki Takehara
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  5. Masami Hayashi
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  6. Daisuke Tachibana
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  7. Hideki Wanibuchi
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  8. Shoji Fukushima
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  9. Masayasu Koyama
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  10. Kayo Yoshida
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  11. Takashi Morita
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Corresponding author

Correspondence to Kayo Yoshida.

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Responsible editor: Markus Hecker

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Kitada, K., Kizu, A., Teramura, T. et al. Gene-modified embryonic stem cell test to characterize chemical risks. Environ Sci Pollut Res 22, 18252–18259 (2015). https://doi.org/10.1007/s11356-015-5051-0

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  • DOI: https://doi.org/10.1007/s11356-015-5051-0

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