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Mechanical memory and dosing influence stem cell fate

Abstract

We investigated whether stem cells remember past physical signals and whether these can be exploited to dose cells mechanically. We found that the activation of the Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding domain (TAZ) as well as the pre-osteogenic transcription factor RUNX2 in human mesenchymal stem cells (hMSCs) cultured on soft poly(ethylene glycol) (PEG) hydrogels (Young’s modulus E ~ 2 kPa) depended on previous culture time on stiff tissue culture polystyrene (TCPS; E ~ 3 GPa). In addition, mechanical dosing of hMSCs cultured on initially stiff (E ~ 10 kPa) and then soft (E ~ 2 kPa) phototunable PEG hydrogels resulted in either reversible or—above a threshold mechanical dose—irreversible activation of YAP/TAZ and RUNX2. We also found that increased mechanical dosing on supraphysiologically stiff TCPS biases hMSCs towards osteogenic differentiation. We conclude that stem cells possess mechanical memory—with YAP/TAZ acting as an intracellular mechanical rheostat—that stores information from past physical environments and influences the cells’ fate.

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Figure 1: Mechanical dosing and memory of hMSCs.
Figure 2: Influence of phototunable substrate modulus on YAP and RUNX2 activation.
Figure 3: Reversible and irreversible effects of mechanical dosing on phototunable hydrogels.
Figure 4: Influence of mechanical dosing on differentiation of hMSCs.

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References

  1. Pelham, R. J. & Wang, Y. L. Cell locomotion and focal adhesions are regulated by substrate flexibility. Proc. Natl Acad. Sci. USA 94, 13661–13665 (1997).

    Article  CAS  Google Scholar 

  2. McBeath, R., Pirone, D. M., Nelson, C. M., Bhadriraju, K. & Chen, C. S. Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev. Cell 6, 483–495 (2004).

    Article  CAS  Google Scholar 

  3. Kilian, K. A., Bugarija, B., Lahn, B. T. & Mrksich, M. Geometric cues for directing the differentiation of mesenchymal stem cells. Proc. Natl Acad. Sci. USA 107, 4872–4877 (2010).

    Article  CAS  Google Scholar 

  4. McMurray, R. J. et al. Nanoscale surfaces for the long-term maintenance of mesenchymal stem cell phenotype and multipotency. Nature Mater. 10, 637–644 (2011).

    Article  CAS  Google Scholar 

  5. Guvendiren, M. & Burdick, J. A. Stiffening hydrogels to probe short- and long-term cellular responses to dynamic mechanics. Nature Commun. 3, 792 (2012).

    Article  Google Scholar 

  6. Trappmann, B. et al. Extracellular-matrix tethering regulates stem-cell fate. Nature Mater. 11, 642–649 (2012).

    Article  CAS  Google Scholar 

  7. Khetan, S. et al. Degradation-mediated cellular traction directs stem cell fate in covalently crosslinked three-dimensional hydrogels. Nature Mater. 12, 458–465 (2013).

    Article  CAS  Google Scholar 

  8. Engler, A. J., Sen, S., Sweeney, H. L. & Discher, D. E. Matrix elasticity directs stem cell lineage specification. Cell 126, 677–689 (2006).

    Article  CAS  Google Scholar 

  9. Discher, D. E., Mooney, D. J. & Zandstra, P. W. Growth factors, matrices, and forces combine and control stem cells. Science 324, 1673–1677 (2009).

    Article  CAS  Google Scholar 

  10. Guilak, F. et al. Control of stem cell fate by physical interactions with the extracellular matrix. Cell Stem Cell 5, 17–26 (2009).

    Article  CAS  Google Scholar 

  11. Watt, F. M., Jordan, P. W. & O’Neill, C. H. Cell shape controls terminal differentiation of human epidermal-keratinocytes. Proc. Natl Acad. Sci. USA 85, 5576–5580 (1988).

    Article  CAS  Google Scholar 

  12. Ingber, D. E. Fibronectin controls capillary endothelial-cell growth by modulating cell shape. Proc. Natl Acad. Sci. USA 87, 3579–3583 (1990).

    Article  CAS  Google Scholar 

  13. Chen, C. S., Mrksich, M., Huang, S., Whitesides, G. M. & Ingber, D. E. Geometric control of cell life and death. Science 276, 1425–1428 (1997).

    Article  CAS  Google Scholar 

  14. Gilbert, P. M. et al. Substrate elasticity regulates skeletal muscle stem cell self-renewal in culture. Science 329, 1078–1081 (2010).

    Article  CAS  Google Scholar 

  15. Dupont, S. et al. Role of YAP/TAZ in mechanotransduction. Nature 474, 179–183 (2011).

    Article  CAS  Google Scholar 

  16. Halder, G., Dupont, S. & Piccolo, S. Transduction of mechanical and cytoskeletal cues by YAP and TAZ. Nat. Rev. Mol. Cell Biol. 13, 591–600 (2012).

    Article  CAS  Google Scholar 

  17. Maloney, J. M. et al. Mesenchymal stem cell mechanics from the attached to the suspended state. Biophys. J. 99, 2479–2487 (2010).

    Article  CAS  Google Scholar 

  18. Kloxin, A. M., Kasko, A. M., Salinas, C. N. & Anseth, K. S. Photodegradable hydrogels for dynamic tuning of physical and chemical properties. Science 324, 59–63 (2009).

    Article  CAS  Google Scholar 

  19. Kloxin, A. M., Tibbitt, M. W. & Anseth, K. S. Synthesis of photodegradable hydrogels as dynamically tunable cell culture platforms. Nature Protocols 5, 1867–1887 (2010).

    Article  CAS  Google Scholar 

  20. Kloxin, A. M., Tibbitt, M. W., Kasko, A. M., Fairbairn, J. F. & Anseth, K. S. Tunable hydrogels for external manipulation of cellular microenvironments through controlled photodegradation. Adv. Mater. 22, 61–66 (2010).

    Article  CAS  Google Scholar 

  21. Tibbitt, M. W., Kloxin, A. M. & Anseth, K. S. Modeling controlled photodegradation in optically thick hydrogels. J. Polym. Sci. Pol. Chem. 51, 1899–1911 (2013).

    Article  CAS  Google Scholar 

  22. Tibbitt, M. W., Kloxin, A. M., Sawicki, L. A. & Anseth, K. S. Mechanical properties and degradation of chain and step-polymerized photodegradable hydrogels. Macromolecules 46, 2785–2792 (2013).

    Article  CAS  Google Scholar 

  23. Wong, D. Y., Griffin, D. R., Reed, J. & Kasko, A. M. Photodegradable hydrogels to generate positive and negative features over multiple length scales. Macromolecules 43, 2824–2831 (2010).

    Article  CAS  Google Scholar 

  24. Kloxin, A. M., Benton, J. A. & Anseth, K. S. In situ elasticity modulation with dynamic substrates to direct cell phenotype. Biomaterials 31, 1–8 (2010).

    Article  CAS  Google Scholar 

  25. Tibbitt, M. W., Kloxin, A. M., Dyamenahalli, K. U. & Anseth, K. S. Controlled two-photon photodegradation of PEG hydrogels to study and manipulate subcellular interactions on soft materials. Soft Matter 6, 5100–5108 (2010).

    Article  CAS  Google Scholar 

  26. Wang, H., Haeger, S. M., Kloxin, A. M., Leinwand, L. A. & Anseth, K. S. Redirecting valvular myofibroblasts into dormant fibroblasts through light-mediated reduction in substrate modulus. PloS ONE 7, e39969 (2012).

    Article  CAS  Google Scholar 

  27. Frey, M. T. & Wang, Y. L. A photo-modulatable material for probing cellular responses to substrate rigidity. Soft Matter 5, 1918–1924 (2009).

    Article  CAS  Google Scholar 

  28. Wang, H., Tibbitt, M. W., Langer, S. J., Leinwand, L. A. & Anseth, K. S. Hydrogels preserve native phenotypes of valvular fibroblasts through an elasticity-regulated pi3k/akt pathway. Proc. Natl Acad. Sci. USA 110, 19336–19341 (2013).

    Article  CAS  Google Scholar 

  29. Pittenger, M. F. et al. Multilineage potential of adult human mesenchymal stem cells. Science 284, 143–147 (1999).

    Article  CAS  Google Scholar 

  30. Shin, J. W. et al. Mechanobiology of bone marrow stem cells: From myosin-II forces to compliance of matrix and nucleus in cell forms and fates. Differentiation 86, 77–86 (2013).

    Article  CAS  Google Scholar 

  31. Mariner, P. D., Johannesen, E. & Anseth, K. S. Manipulation of miRNA activity accelerates osteogenic differentiation of hMSCs in engineered 3d scaffolds. J. Tissue Eng. Regen. Med. 6, 314–324 (2012).

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by the National Science Foundation (DMR 1006711; K.S.A.), the National Institutes of Health (1R21 AR057904, R01 DE016523), the Howard Hughes Medical Institute (K.S.A.), the Teets Family Endowed Doctoral Fellowship (M.W.T.), and the Molecular Biophysics Training Grant from the National Institutes of Health (T32 GM-065103; M.W.T.). We would like to thank R. Tjian and I. Grubisic for helpful discussions on the work as well as E. A. Appel and T. A. Tauer for advice on figure preparation.

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M.W.T., C.Y. and K.S.A. conceived the ideas and designed the experiments. C.Y., L.B. and M.W.T. conducted the experiments and analysed the data. M.W.T., C.Y., L.B. and K.S.A. interpreted the data and wrote the manuscript.

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Yang, C., Tibbitt, M., Basta, L. et al. Mechanical memory and dosing influence stem cell fate. Nature Mater 13, 645–652 (2014). https://doi.org/10.1038/nmat3889

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