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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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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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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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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DOI: https://doi.org/10.1038/nmat3339
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