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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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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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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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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DOI: https://doi.org/10.1038/nmat4051
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