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Interplay of matrix stiffness and protein tethering in stem cell differentiation

Nature Materials volume 13, pages 979–987 (2014)Cite this article

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Abstract

Stem cells regulate their fate by binding to, and contracting against, the extracellular matrix. Recently, it has been proposed that in addition to matrix stiffness and ligand type, the degree of coupling of fibrous protein to the surface of the underlying substrate, that is, tethering and matrix porosity, also regulates stem cell differentiation. By modulating substrate porosity without altering stiffness in polyacrylamide gels, we show that varying substrate porosity did not significantly change protein tethering, substrate deformations, or the osteogenic and adipogenic differentiation of human adipose-derived stromal cells and marrow-derived mesenchymal stromal cells. Varying protein–substrate linker density up to 50-fold changed tethering, but did not affect osteogenesis, adipogenesis, surface–protein unfolding or underlying substrate deformations. Differentiation was also unaffected by the absence of protein tethering. Our findings imply that the stiffness of planar matrices regulates stem cell differentiation independently of protein tethering and porosity.

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Figure 1: Influence of substrate porosity on ASC differentiation.
Figure 2: Influence of protein tethering on ASC differentiation.
Figure 3: Direct incorporation of a short adhesive peptide to the PA substrate.
Figure 4: Cells sense by contracting against their substrates.
Figure 5: Atomic force spectrography analysis of ligand-coated PDMS substrates.

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Acknowledgements

The authors thank M. Joens, J. Kasuboski and J. Fitzpatrick at the Waitt Advanced Biophotonics Center at the Salk Institute (supported by NCI P30 Cancer Center Support Grant CA014195-40 and NINDS P30 Neuroscience Center Core Grant NS072031-03A1 and by the W. M. Keck Foundation) for technical assistance with microscopy, and W. Murphy (University of Wisconsin), T. McDevitt (Georgia Tech) and J. C. del Alamo (UC San Diego) for helpful conversations. The National Institutes of Health (DP02OD006460 to A.J.E.), the Human Frontiers Science Program (RGY0064/2010 to A.J.E.), the National Science Foundation Graduate Research Fellowship Program (to J.H.W., L.G.V. and H.T-W.), the Siebel Scholars Program, and the Achievement Rewards for College Scientists (to L.G.V.) supported this work.

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Author notes

  1. Yu Suk Choi

    Present address: Present address: Kolling Institute of Medical Research, Royal North Shore Hospital, The University of Sydney, Sydney, New South Wales 2065, Australia.,

  2. Jessica H. Wen and Ludovic G. Vincent: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Bioengineering, University of California, San Diego, La Jolla, California 92093, USA

    Jessica H. Wen, Ludovic G. Vincent, Alexander Fuhrmann, Yu Suk Choi, Hermes Taylor-Weiner & Adam J. Engler

  2. Department of Nanoengineering, University of California, San Diego, La Jolla, California 92093, USA

    Kolin C. Hribar & Shaochen Chen

  3. Sanford Consortium for Regenerative Medicine, La Jolla, California 92037, USA

    Adam J. Engler

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Contributions

All authors contributed to the design of experiments. Y.S.C., K.C.H. and S.C. characterized the hydrogel substrates. H.T-W. characterized and performed experiments with the FRET probe. J.H.W., L.G.V. and A.F. conducted all other experiments and performed the data analysis. J.H.W., L.G.V. and A.J.E. wrote the manuscript.

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Correspondence to Adam J. Engler.

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The authors declare no competing financial interests.

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Wen, J., Vincent, L., Fuhrmann, A. et al. Interplay of matrix stiffness and protein tethering in stem cell differentiation. Nature Mater 13, 979–987 (2014). https://doi.org/10.1038/nmat4051

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