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Defined three-dimensional microenvironments boost induction of pluripotency

Nature Materials volume 15, pages 344–352 (2016)Cite this article

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Abstract

Since the discovery of induced pluripotent stem cells (iPSCs), numerous approaches have been explored to improve the original protocol, which is based on a two-dimensional (2D) cell-culture system. Surprisingly, nothing is known about the effect of a more biologically faithful 3D environment on somatic-cell reprogramming. Here, we report a systematic analysis of how reprogramming of somatic cells occurs within engineered 3D extracellular matrices. By modulating microenvironmental stiffness, degradability and biochemical composition, we have identified a previously unknown role for biophysical effectors in the promotion of iPSC generation. We find that the physical cell confinement imposed by the 3D microenvironment boosts reprogramming through an accelerated mesenchymal-to-epithelial transition and increased epigenetic remodelling. We conclude that 3D microenvironmental signals act synergistically with reprogramming transcription factors to increase somatic plasticity.

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Figure 1: 3D PEG hydrogel cultures maintain ESC pluripotency.
Figure 2: Generation of 3DiPSCs.
Figure 3: 3D culture accelerates reprogramming and increases iPSC generation efficiency.
Figure 4: Optimization of iPSC reprogramming efficiency by modulation of 3D microenvironment.
Figure 5: 3D reprogramming accelerates MET and enhances epigenetic plasticity.
Figure 6: 3D generation of human iPSCs.

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Acknowledgements

We thank M. Snyder and M. Knobloch for critical reading of the manuscript, A. Negro for polymer batch testing, D. Trono for providing Oct4–GFP and OKSM-doxy-inducible mice, M. Friedli for providing SFFV-OKSM lentiviral vector and for helpful discussion, M. Pluchinotta and P. Manti for the anti-E-Cad and anti-SMA antibodies, the EPFL Transgenic Core Facilities for generating chimaeric mice, and the EPFL Histology Core Facility for performing teratoma histology. This work was financially supported by the EU framework 7 HEALTH research programme PluriMes (http://www.plurimes.eu), the SystemsX.ch RTD project StoNets, an ERC grant (StG_311422) and a Swiss National Science Foundation Singergia grant (CRSII3_147684).

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Author notes

  1. Massimiliano Caiazzo and Yuya Okawa: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Stem Cell Bioengineering, Institute of Bioengineering, School of Life Sciences (SV) and School of Engineering (STI), Ecole Polytechnique Fédérale de Lausanne (EPFL), 1015 Lausanne, Switzerland

    Massimiliano Caiazzo, Yuya Okawa, Adrian Ranga, Yoji Tabata & Matthias P. Lutolf

  2. School of Life Sciences, Core Facility PTECH, EPFL, 1015 Lausanne, Switzerland

    Alessandra Piersigilli

  3. Institute of Chemical Sciences and Engineering, School of Basic Science (SB), EPFL, 1015 Lausanne, Switzerland

    Matthias P. Lutolf

Authors
  1. Massimiliano Caiazzo
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  2. Yuya Okawa
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Contributions

M.P.L., Y.O. and M.C. designed the experiments and analysed the data. Y.O. and M.C. performed most of the experiments and statistical analyses. M.P.L. and M.C. wrote the manuscript. A.R. performed and analysed the HTS experiment and contributed to manuscript writing. Y.T. fabricated the microgroove platform. A.P. analysed teratoma assays.

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Correspondence to Matthias P. Lutolf.

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The authors declare no competing financial interests.

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Caiazzo, M., Okawa, Y., Ranga, A. et al. Defined three-dimensional microenvironments boost induction of pluripotency. Nature Mater 15, 344–352 (2016). https://doi.org/10.1038/nmat4536

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