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Inhibition of TGFβ Signaling Promotes Ground State Pluripotency

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Abstract

Embryonic stem (ES) cells are considered to exist in a ground state if shielded from differentiation triggers. Here we show that FGF4 and TGFβ signaling pathway inhibitors, designated R2i, not only provide the ground state pluripotency in production and maintenance of naïve ES cells from blastocysts of different mouse strains, but also maintain ES cells with higher genomic integrity following long-term cultivation compared with the chemical inhibition of the FGF4 and GSK3 pathways, known as 2i. Global transcriptome analysis of the ES cells highlights augmented BMP4 signaling pathway. The crucial role of the BMP4 pathway in maintaining the R2i ground state pluripotency is demonstrated by BMP4 receptor suppression, resulting in differentiation and cell death. In conclusion, by inhibiting TGFβ and FGF signaling pathways, we introduce a novel defined approach to efficiently establish the ground state pluripotency.

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Acknowledgments

We thank the members of the Department of Stem Cells and Developmental Biology labs for their helpful suggestions and critical reading of the manuscript. We thank Behrouz Asgari for chimera formation and germline transmission. This study was funded by grants provided from Royan Institute and Iranian Council of Stem Cell Technology and the Iran National Science Foundation (INSF).

Author Contributions

S. H., M. T., H. R. S. and H. B. designed all experiments and wrote the manuscript. S. H. and S. M. performed cell culture. M. T. and A. S. performed real-time PCR analysis. A. F. and M. P. operated in vivo experiments. N. M. and H. G. performed karyotype analysis. S. M. and G. S. H. performed western blot analysis. M. T., A. S., M. S., B. G. and M. J. A. designed and interpreted microarray analysis. G. H. S., S. M. and D. S. contributed to the overall design and writing of the article.

Conflict of Interest

The authors indicate no potential conflicts of interest.

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Authors and Affiliations

  1. Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, PO Box 19395–4644, Tehran, Iran

    Seyedeh-Nafiseh Hassani, Mehdi Totonchi, Ali Sharifi-Zarchi, Sepideh Mollamohammadi, Mohammad Pakzad, Sharif Moradi, Azam Samadian, Ali Farrokhi, Davood Sabour & Hossein Baharvand

  2. Department of Developmental Biology, University of Science and Culture, ACECR, Tehran, Iran

    Seyedeh-Nafiseh Hassani, Mehdi Totonchi & Hossein Baharvand

  3. Department of Genetics at Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

    Mehdi Totonchi, Najmehsadat Masoudi & Hamid Gourabi

  4. Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran

    Ali Sharifi-Zarchi

  5. National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

    Mehdi Sadeghi

  6. Department of Molecular Systems Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

    Shahab Mirshahvaladi & Ghasem Hosseini Salekdeh

  7. Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstrasse 20, 48149, Münster, Germany

    Boris Greber, Marcos J. Araúzo-Bravo, Davood Sabour & Hans R. Schöler

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  1. Seyedeh-Nafiseh Hassani
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  2. Mehdi Totonchi
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Corresponding author

Correspondence to Hossein Baharvand.

Additional information

Seyedeh-Nafiseh Hassani and Mehdi Totonchi contributed equally in this work.

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Hassani, SN., Totonchi, M., Sharifi-Zarchi, A. et al. Inhibition of TGFβ Signaling Promotes Ground State Pluripotency. Stem Cell Rev and Rep 10, 16–30 (2014). https://doi.org/10.1007/s12015-013-9473-0

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