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Extracellular-matrix tethering regulates stem-cell fate

Nature Materials volume 11, pages 642–649 (2012)Cite this article

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A Corrigendum to this article was published on 24 July 2012

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Abstract

To investigate how substrate properties influence stem-cell fate, we cultured single human epidermal stem cells on polydimethylsiloxane (PDMS) and polyacrylamide (PAAm) hydrogel surfaces, 0.1 kPa–2.3 MPa in stiffness, with a covalently attached collagen coating. Cell spreading and differentiation were unaffected by polydimethylsiloxane stiffness. However, cells on polyacrylamide of low elastic modulus (0.5 kPa) could not form stable focal adhesions and differentiated as a result of decreased activation of the extracellular-signal-related kinase (ERK)/mitogen-activated protein kinase (MAPK) signalling pathway. The differentiation of human mesenchymal stem cells was also unaffected by PDMS stiffness but regulated by the elastic modulus of PAAm. Dextran penetration measurements indicated that polyacrylamide substrates of low elastic modulus were more porous than stiff substrates, suggesting that the collagen anchoring points would be further apart. We then changed collagen crosslink concentration and used hydrogel–nanoparticle substrates to vary anchoring distance at constant substrate stiffness. Lower collagen anchoring density resulted in increased differentiation. We conclude that stem cells exert a mechanical force on collagen fibres and gauge the feedback to make cell-fate decisions.

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Figure 1: Influence of substrate elasticity on the spreading and differentiation of human keratinocytes.
Figure 2: Differentiation on soft PAAm substrates correlates with reduced SRF activity, reduced focal adhesion formation and reduced ERK/MAPK signalling.
Figure 3: Influence of substrate elasticity on spreading and differentiation of hMSCs.
Figure 4: Collagen attachment varies on polyacrylamide hydrogels of different stiffnesses.

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Change history

  • 03 July 2012

    In the version of this Article originally published, in Fig. 4f, the two red-stained fluorescence microscopy images were reversed; this has now been corrected in the HTML and PDF versions.

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Acknowledgements

This work was supported by the Wellcome Trust (F.M.W.), the Medical Research Council (F.M.W.), the European Union FP7 (F.M.W.) and a pump-priming grant from Cancer Research UK (W.T.S.H.). B.T. is the recipient of a Gates Cambridge Scholarship. We thank R. Treisman for providing reagents. We are grateful to J. Skepper, A. Bahnweg, J. Shapiro and the core facilities of the Wellcome Trust Centre for Stem Cell Research for technical assistance. We would like to thank Y. Schoen for preparation of the nanopatterned hydrogels. J. Burdick, C. Chen, B. Baker, F. Oceguera-Yanez and K. Kretzschmar are thanked for discussions and advice. We acknowledge financial support from the Max Planck Society (J.P.S. and H.B.), a European Research Council Advanced Grant (V.V.) and Swiss Federal Institute of Technology Zurich (B.L., V.V.).

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Authors and Affiliations

  1. Department of Chemistry, Melville Laboratory for Polymer Synthesis, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK

    Britta Trappmann, Julien E. Gautrot & Wilhelm T. S. Huck

  2. Wellcome Trust Centre for Stem Cell Research, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK

    Julien E. Gautrot, John T. Connelly & Fiona M. Watt

  3. Blizard Institute of Cell and Molecular Science School of Medicine and Dentistry, Queen Mary, University of London, Mile End Road, London E1 4NS, UK

    John T. Connelly

  4. Department of Engineering, University of Cambridge, Trumpington Street, Cambridge CB2 1PZ, UK

    Daniel G. T. Strange & Michelle L. Oyen

  5. Laboratory of Physical Chemistry and Colloid Science, Dreijenplein 6, 6703 HB Wageningen, The Netherlands

    Yuan Li & Martien A. Cohen Stuart

  6. Department of New Materials and Biosystems, Max Planck Institute for Intelligent Systems, Heisenbergstrasse 3, 70569 Stuttgart, Germany

    Heike Boehm & Joachim P. Spatz

  7. Department of Biophysical Chemistry, University of Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, Germany

    Heike Boehm & Joachim P. Spatz

  8. Department of Health Sciences and Technology, ETH Zurich, Wolfgang-Pauli-Strasse 10, CH-8093 Zurich, HCI F443, Switzerland

    Bojun Li & Viola Vogel

  9. CRUK Cambridge Research Institute, Robinson Way, Cambridge CB2 0RE, UK

    Fiona M. Watt

  10. Radboud University Nijmegen, Institute for Molecules and Materials, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands

    Wilhelm T. S. Huck

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  1. Britta Trappmann
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Contributions

The experiments were designed by B.T., J.E.G., F.M.W. and W.T.S.H., and carried out by B.T. J.E.G. acquired time-lapse recordings, carried out fibronectin experiments and helped with collagen immunofluorescence. M.L.O., B.T. and D.G.T.S. carried out the mechanical characterization of the materials. Y.L. and M.A.C.S. determined the pore sizes using fluorescently labelled dextran. H.B. and J.P.S. provided nanoparticle-embedded hydrogels. B.L. and V.V. assisted with hMSC culture and carried out fibronectin strain FRET-sensor experiments. The data were interpreted by B.T., J.E.G., J.T.C., V.V., F.M.W. and W.T.S.H. The manuscript was written by B.T., F.M.W. and W.T.S.H.

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Correspondence to Fiona M. Watt or Wilhelm T. S. Huck.

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Trappmann, B., Gautrot, J., Connelly, J. et al. Extracellular-matrix tethering regulates stem-cell fate. Nature Mater 11, 642–649 (2012). https://doi.org/10.1038/nmat3339

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