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Harnessing traction-mediated manipulation of the cell/matrix interface to control stem-cell fate

Nature Materials volume 9, pages 518–526 (2010)Cite this article

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Abstract

Stem cells sense and respond to the mechanical properties of the extracellular matrix. However, both the extent to which extracellular-matrix mechanics affect stem-cell fate in three-dimensional microenvironments and the underlying biophysical mechanisms are unclear. We demonstrate that the commitment of mesenchymal stem-cell populations changes in response to the rigidity of three-dimensional microenvironments, with osteogenesis occurring predominantly at 11–30 kPa. In contrast to previous two-dimensional work, however, cell fate was not correlated with morphology. Instead, matrix stiffness regulated integrin binding as well as reorganization of adhesion ligands on the nanoscale, both of which were traction dependent and correlated with osteogenic commitment of mesenchymal stem-cell populations. These findings suggest that cells interpret changes in the physical properties of adhesion substrates as changes in adhesion-ligand presentation, and that cells themselves can be harnessed as tools to mechanically process materials into structures that feed back to manipulate their fate.

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Figure 1: Matrix compliance alters MSC fate in 3D matrix culture.
Figure 2: Cell and nuclear morphology are not strongly correlated to mechanics of 3D matrices.
Figure 3: Mechanically controlled α5-integrin–RGD bond formation correlates with stem-cell osteogenic lineage in three dimensions.
Figure 4: Cell–RGD bond formation shows a biphasic dependence on matrix stiffness.
Figure 5: Cell-traction-mediated reorganization of ligands presented by synthetic ECM.
Figure 6: Long-term regulation of osteogenic commitment and role of specific integrins in stem-cell fate in 3D matrices.

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Acknowledgements

We thank R. F. Horwitz for providing the human α5-integrin–enhanced GFP plasmid, and B. Tilton for carrying out FACs sorting of GFP-expressing mMSCs at the Harvard FAS Center for Systems Biology. We also thank E. Gatzogiannis for assisting with confocal microscopy at the Harvard University Center for Nanoscale Systems (CNS). The monoclonal antibody against osteopontin (MPIIIB101) developed by M. Solursh and A. Franzen was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Department of Biology, Iowa City, IA. We acknowledge R. Schmidt and K. E. Healy (University of California, Berkeley) for advice regarding nuclear morphology measurements, A. Putnam (University of Michigan), H. J. Kong, C. Fischbach and S. Hsiong for discussions, and H. Vandenburgh (Brown University) and Z. Suo for critical reading of the manuscript. We also acknowledge funding from NIH (R37 DE013033) and from the NIH ‘Systems-Based Consortium for Organ Design and Engineering’ training grant (J.R-F., 1TL1EB008540-01, NIBIB), a Harvard Presidential Fellowship (P.R.A.), an NSF Graduate Research Fellowship (N.H.), the Harvard College Research Program (A.S.M.), an EMBO Long-Term Fellowship ALTF 42-2008 (D.S.) and the Wyss Institute for Biologically Inspired Engineering (S.A.B.).

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  1. Omar A. Ali

    Present address: Present address: InCytu, Inc, Lincoln, Rhode Island 02865, USA,

Authors and Affiliations

  1. School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts 02138, USA

    Nathaniel Huebsch, Praveen R. Arany, Angelo S. Mao, Dmitry Shvartsman, Omar A. Ali, Sidi A. Bencherif, José Rivera-Feliciano & David J. Mooney

  2. Harvard-MIT Division of Health Sciences and Technology, Cambridge, Massachusetts 02138, USA

    Nathaniel Huebsch

  3. Wyss Institute for Biologically Inspired Engineering, Cambridge, Massachusetts 02138, USA

    Nathaniel Huebsch, Praveen R. Arany, Dmitry Shvartsman, Sidi A. Bencherif & David J. Mooney

  4. Programs in Oral and Maxillofacial Pathology, Leder Human Biology and Translational Medicine and Biological Sciences in Dental Medicine, Harvard School of Dental Medicine & Brigham and Women’s Hospital, Boston, Massachusetts 02115, USA

    Praveen R. Arany

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Contributions

The experiments were designed by N.H., P.R.A. and D.J.M. and carried out by N.H., P.R.A., A.S.M., D.S. and O.A.A. N.H., S.A.B. and J.R-F. provided new reagents/analytical tools. The manuscript was written by N.H. and D.J.M. The principal investigator is D.J.M.

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Correspondence to David J. Mooney.

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Huebsch, N., Arany, P., Mao, A. et al. Harnessing traction-mediated manipulation of the cell/matrix interface to control stem-cell fate. Nature Mater 9, 518–526 (2010). https://doi.org/10.1038/nmat2732

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