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Thalidomide induces apoptosis in undifferentiated human induced pluripotent stem cells

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Abstract

Thalidomide, which was formerly available commercially to control the symptoms of morning sickness, is a strong teratogen that causes fetal abnormalities. However, the mechanism of thalidomide teratogenicity is not fully understood; thalidomide toxicity is not apparent in rodents, and the use of human embryos is ethically and technically untenable. In this study, we designed an experimental system featuring human-induced pluripotent stem cells (hiPSCs) to investigate the effects of thalidomide. These cells exhibit the same characteristics as those of epiblasts originating from implanted fertilized ova, which give rise to the fetus. Therefore, theoretically, thalidomide exposure during hiPSC differentiation is equivalent to that in the human fetus. We examined the effects of thalidomide on undifferentiated hiPSCs and early-differentiated hiPSCs cultured in media containing bone morphogenetic protein-4, which correspond, respectively, to epiblast (future fetus) and trophoblast (future extra-embryonic tissue). We found that only the number of undifferentiated cells was reduced. In undifferentiated cells, application of thalidomide increased the number of apoptotic and dead cells at day 2 but not day 4. Application of thalidomide did not affect the cell cycle. Furthermore, immunostaining and flow cytometric analysis revealed that thalidomide exposure had no effect on the expression of specific markers of undifferentiated and early trophectodermal differentiated cells. These results suggest that the effect of thalidomide was successfully detected in our experimental system and that thalidomide eliminated a subpopulation of undifferentiated hiPSCs. This study may help to elucidate the mechanisms underlying thalidomide teratogenicity and reveal potential strategies for safely prescribing this drug to pregnant women.

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Acknowledgments

This work was supported in part by grants from the Ministry of Health, Labour, and Welfare of Japan, the Japan Agency for Medical Research and Development (AMED) (to K.O.), and the Foundation for Applied Research and Technological Uniqueness at Nagaoka University of Technology (to S.T.). The funding bodies had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. Department of Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata, 940-2188, Japan

    Saoko Tachikawa & Kiyoshi Ohnuma

  2. Division of Stem Cell Therapy, Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, University of Tokyo, Tokyo, 108-8639, Japan

    Toshinobu Nishimura & Hiromitsu Nakauchi

  3. Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, 94305, USA

    Hiromitsu Nakauchi

  4. Department of Science of Technology Innovation, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata, 940-2188, Japan

    Kiyoshi Ohnuma

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  1. Saoko Tachikawa
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  2. Toshinobu Nishimura
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  3. Hiromitsu Nakauchi
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  4. Kiyoshi Ohnuma
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Contributions

S.T. and K.O. designed the project. S.T. performed all experiments. T.N. and H.N. provided a hiPS clone and helpful feedback on the manuscript. S.T. and K.O. wrote the manuscript and all authors reviewed the manuscript.

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Correspondence to Kiyoshi Ohnuma.

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The authors declare that they have no competing financial interests.

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Editor: Tetsuji Okamoto

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Tachikawa, S., Nishimura, T., Nakauchi, H. et al. Thalidomide induces apoptosis in undifferentiated human induced pluripotent stem cells. In Vitro Cell.Dev.Biol.-Animal 53, 841–851 (2017). https://doi.org/10.1007/s11626-017-0192-8

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  • DOI: https://doi.org/10.1007/s11626-017-0192-8

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