Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Substrate area confinement is a key determinant of cell velocity in collective migration

Nature Physics volume 15, pages 858–866 (2019)Cite this article

Subjects

Abstract

Collective cell migration is fundamental throughout development, during wound healing and in many diseases. Although much effort has focused on cell–cell junctions, a role for physical confinement in collective cell migration remains unclear. Here, we used adhesive microstripes of varying widths to mimic the spatial confinement experienced by follower cells within epithelial tissues. Our results reveal that the substrate area confinement is sufficient to modulate the three-dimensional cellular morphology without the need for intercellular adhesive cues. Our findings show a direct correlation between the migration velocity of confined cells and their cell–substrate adhesive area. Closer examination revealed that adhesive area confinement reduces lamellipodial protrusive forces, decreases the number of focal complexes at the leading edge and prevents the maturation of focal adhesions at the trailing edge, together leading to less effective forward propelling forces. The release of follower confinement required for the emergence of leader cells is associated with a threefold increase in contractile stress and a tenfold increase in protrusive forces, together providing a sufficient stress to generate highly motile mesenchymal cells. These findings demonstrate that epithelial confinement alone can induce follower-like behaviours and identify substrate adhesive area confinement as a key determinant of cell velocity in collective migration.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Epithelial cells are confined within epithelial tissues.
Fig. 2: Cell morphologies and migration velocities are regulated by the lateral confinement.
Fig. 3: 2D confinement modulates the 3D cellular morphology.
Fig. 4: Thicker lamellipodia exert less protrusive forces.
Fig. 5: Escaping a tissue requires the reorganization of the actin cytoskeleton.
Fig. 6: Migration velocity is correlated to cell–substrate adhesive area.

Similar content being viewed by others

Data availability

The data that support the plots within this paper and other findings of this study are available from the corresponding author upon request.

References

  1. Keller, R. Shaping the vertebrate body plan by polarized embryonic cell movements. Science 298, 1950–1954 (2002).

    Article  ADS  Google Scholar 

  2. Farooqui, R. & Fenteany, G. Multiple rows of cells behind an epithelial wound edge extend cryptic lamellipodia to collectively drive cell-sheet movement. J. Cell Sci. 118, 51–63 (2004).

    Article  Google Scholar 

  3. Friedl, P. & Gilmour, D. Collective cell migration in morphogenesis, regeneration and cancer. Nat. Rev. Mol. Cell Biol. 10, 445–457 (2009).

    Article  Google Scholar 

  4. Ilina, O. & Friedl, P. Mechanisms of collective cell migration at a glance. J. Cell Sci. 122, 3203–3208 (2009).

    Article  Google Scholar 

  5. Ridley, A. J. Cell migration: integrating signals from front to back. Science 302, 1704–1709 (2003).

    Article  ADS  Google Scholar 

  6. Murrell, M., Oakes, P. W., Lenz, M. & Gardel, M. L. Forcing cells into shape: the mechanics of actomyosin contractility. Nat. Rev. Mol. Cell Biol. 16, 486–498 (2015).

    Article  Google Scholar 

  7. Mayor, R. & Etienne-Manneville, S. The front and rear of collective cell migration. Nat. Rev. Mol. Cell Biol. 17, 97–109 (2016).

    Article  Google Scholar 

  8. Friedl, P. & Alexander, S. Cancer invasion and the microenvironment: plasticity and reciprocity. Cell 147, 992–1009 (2011).

    Article  Google Scholar 

  9. Wong, I. Y. et al. Collective and individual migration following the epithelial–mesenchymal transition. Nat. Mater. 13, 1063–1071 (2014).

    Article  ADS  Google Scholar 

  10. Vedula, S. R. K. et al. Emerging modes of collective cell migration induced by geometrical constraints. Proc. Natl Acad. Sci. USA 109, 12974–12979 (2012).

    Article  ADS  Google Scholar 

  11. Abercrombie, M. & Heaysman, J. E. Observations on the social behaviour of cells in tissue culture. I. Speed of movement of chick heart fibroblasts in relation to their mutual contacts. Exp. Cell Res. 5, 111–131 (1953).

    Article  Google Scholar 

  12. Stoker, M. G. P. & Rubin, H. Density dependent inhibition of cell growth in culture. Nature 215, 171–172 (1967).

    Article  ADS  Google Scholar 

  13. Angelini, T. E. et al. Glass-like dynamics of collective cell migration. Proc. Natl Acad. Sci. USA 108, 4714–4719 (2011).

    Article  ADS  Google Scholar 

  14. Garcia, S. et al. Physics of active jamming during collective cellular motion in a monolayer. Proc. Natl Acad. Sci. USA 112, 15314–15319 (2015).

    Article  ADS  Google Scholar 

  15. Keren, K. et al. Mechanism of shape determination in motile cells. Nature 453, 475–480 (2008).

    Article  ADS  Google Scholar 

  16. Lacayo, C. I. et al. Emergence of large-scale cell morphology and movement from local actin filament growth dynamics. PLoS Biol. 5, e233 (2007).

    Article  Google Scholar 

  17. Fournier, M. F., Sauser, R., Ambrosi, D., Meister, J.-J. & Verkhovsky, A. B. Force transmission in migrating cells. J. Cell Biol. 188, 287–297 (2010).

    Article  Google Scholar 

  18. Riaz, M., Versaevel, M., Mohammed, D., Glinel, K. & Gabriele, S. Persistence of fan-shaped keratocytes is a matrix-rigidity-dependent mechanism that requires α5β1 integrin engagement. Sci. Rep. 6, 34141 (2016).

    Article  ADS  Google Scholar 

  19. Verkhovsky, A. B. Orientational order of the lamellipodial actin network as demonstrated in living motile cells. Mol. Biol. Cell 14, 4667–4675 (2003).

    Article  Google Scholar 

  20. Ofer, N., Mogilner, A. & Keren, K. Actin disassembly clock determines shape and speed of lamellipodial fragments. Proc. Natl Acad. Sci. USA 108, 20394–20399 (2011).

    Article  ADS  Google Scholar 

  21. Prass, M., Jacobson, K., Mogilner, A. & Radmacher, M. Direct measurement of the lamellipodial protrusive force in a migrating cell. J. Cell Biol. 174, 767–772 (2006).

    Article  Google Scholar 

  22. Mogilner, A. & Oster, G. Cell motility driven by actin polymerization. Biophys. J. 71, 3030–3045 (1996).

    Article  ADS  Google Scholar 

  23. Borm, B., Requardt, R. P., Herzog, V. & Kirfel, G. Membrane ruffles in cell migration: indicators of inefficient lamellipodia adhesion and compartments of actin filament reorganization. Exp. Cell Res. 302, 83–95 (2005).

    Article  Google Scholar 

  24. Rolli, C. G. et al. Switchable adhesive substrates: revealing geometry dependence in collective cell behavior. Biomaterials 33, 2409–2418 (2012).

    Article  Google Scholar 

  25. Trepat, X., Chen, Z. & Jacobson, K. Cell migration. Compr. Physiol. 2, 2369–2392 (2012).

    Google Scholar 

  26. Lantoine, J. et al. Matrix stiffness modulates formation and activity of neuronal networks of controlled architectures. Biomaterials 89, 14–24 (2016).

    Article  Google Scholar 

  27. Versaevel, M., Riaz, M., Grevesse, T. & Gabriele, S. Cell confinement: putting the squeeze on the nucleus. Soft Matter 9, 6665–6676 (2013).

    Article  ADS  Google Scholar 

  28. Versaevel, M., Grevesse, T., Riaz, M., Lantoine, J. & Gabriele, S. Micropatterning hydroxy-PAAm hydrogels and Sylgard 184 silicone elastomers with tunable elastic moduli. Methods Cell Biol. 121, 33–48 (2014).

    Article  Google Scholar 

  29. Coppée, S., Gabriele, S., Jonas, A. M., Jestin, J. & Damman, P. Influence of chain interdiffusion between immiscible polymers on dewetting dynamics. Soft Matter 21, 9951–9955 (2011).

    Article  Google Scholar 

  30. Grevesse, T., Versaevel, M., Circelli, G., Desprez, S. & Gabriele, S. A simple route to functionalize polyacrylamide hydrogels for the independent tuning of mechanotransduction cues. Lab Chip 13, 777–1 (2013).

    Article  Google Scholar 

  31. Grevesse, T., Versaevel, M. & Gabriele, S. Preparation of hydroxy-PAAm hydrogels for decoupling the effects of mechanotransduction cues. J. Vis. Exp. 90, 1–8 (2014).

    Google Scholar 

  32. Versaevel, M., Grevesse, T. & Gabriele, S. Spatial coordination between cell and nuclear shape within micropatterned endothelial cells. Nat. Comm. 3, 671 (2012).

    Article  ADS  Google Scholar 

  33. Butt, H. J. & Jaschke, M. Calculation of thermal noise in atomic force microscopy. Nanotechnology 6, 1–7 (1995).

    Article  ADS  Google Scholar 

  34. Peng, T. et al. BaSiC tool for background and shading correction of optical microscopy images. Nat. Commun. 8, 14836 (2017).

    Article  ADS  Google Scholar 

Download references

Acknowledgements

The authors gratefully acknowledge C.-P. Heisenberg for kindly providing the GFP–UTR zebrafish line, A. Kennard for technical advice for culture of zebrafish embryos and M. Surin for technical support with AFM measurements. The authors thank M. Balland, B. L. Pruitt, M. Sixt, D. Fletcher, J. Theriot and J. Ding for insightful discussions. This work was conducted with the financial support from the Belgian National Fund for Scientific Research (F.R.S.-FNRS, Crédit de Recherches - J009916F) and FEDER Prostem. D.M., J.L., L.A., C.B., E.V. and M.L. are financially supported by FRIA (F.R.S.-FNRS). G.C. is supported by an ERC CoG grant (MolCellTissMech, agreement 647186). K.G. is a Research Associate of the F.R.S.-FNRS.

Author information

Authors and Affiliations

  1. Mechanobiology & Soft Matter Group, Laboratoire Interfaces et Fluides Complexes, Research Institute for Biosciences, University of Mons, Mons, Belgium

    Danahe Mohammed, Eléonore Vercruysse, Marie Versaevel, Joséphine Lantoine, Laura Alaimo, Céline Bruyère, Marine Luciano & Sylvain Gabriele

  2. London Centre for Nanotechnology, University College London, London, UK

    Guillaume Charras

  3. Department of Cell and Developmental Biology, University College London, London, UK

    Guillaume Charras

  4. Institute of Condensed Matter & Nanosciences, Bio & Soft Matter Division, Université catholique de Louvain, Louvain-la-Neuve, Belgium

    Karine Glinel

  5. Adhesion & Inflammation Laboratory, INSERM U1067-CNRS UMR7333, Université Aix-Marseille, Assistance Publique-Hôpitaux de Marseille, Marseille, France

    Geoffrey Delhaye & Olivier Théodoly

Authors
  1. Danahe Mohammed
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Guillaume Charras
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eléonore Vercruysse
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Marie Versaevel
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Joséphine Lantoine
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Laura Alaimo
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Céline Bruyère
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Marine Luciano
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Karine Glinel
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Geoffrey Delhaye
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Olivier Théodoly
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Sylvain Gabriele
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

D.M. and S.G. conceived the project. G.C. and S.G. supervised the project. D.M. and E.V. performed the experiments. C.B. helped with traction force experiments and data analysis. G.D. and O.T. performed PRIMO experiments to create new adhesive micropatterns. K.G. contributed to the silicon wafer microfabrication. M.V., J.L., L.A., C.B. and M.L. contributed resources to the project. D.M., G.C. and S.G. analysed data, wrote the main manuscript text and prepared the figures. All authors contributed to the interpretation of the results, and improved the manuscript and figure presentations.

Corresponding author

Correspondence to Sylvain Gabriele.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Journal peer review information: Nature Physics thanks Kinneret Keren and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information

Supplementary Figs. 1–9.

Reporting Summary

Supplementary Video 1

Time-lapse sequence in DIC mode of the growth of an epithelial tissue from a fish scale deposited on a glass coverslip coated with FN. Total duration: 304 min. The scale bar is 100 μm.

Supplementary Video 2

Time-lapse sequence in DIC mode of the migration of a single keratocyte migrating on a 15-μm-wide microstripe. The FN micropattern is shown in fluorescence mode at t = 5 min. Total duration: 9 min 11 s. The scale bar is 10 μm.

Supplementary Video 3

Time-lapse sequence in DIC mode of the migration of a single keratocyte migrating on a FN micropattern composed of 5 interconnected stripes of different widths: 5, 9, 13, 17 and 21 µm. The total length of the micropattern with variable widths is 320 µm. The total time is 109 min and the scale bar is 10 µm.

Supplementary Video 4

Time-lapse sequence in DIC mode of the cantilever assay to measure protrusive forces. A single keratocyte migrating on a 15-μm-wide microstripe pushed the cantilever. One can observe the cantilever deflection in response to the pushing force exerted by the leading edge of the oncoming cell.

Supplementary Video 5

Time-lapse sequence in DIC mode showing a leader cell that escaped from an epithelial tissue. The red arrow at t = 0 min shows the initial position of the leader cell. The total duration time is 25 min. The scale bar is 25 μm.

Supplementary Video 6

Time-lapse sequence in DIC mode showing a single keratocyte that escaped from a narrow microstripe. The escape process was reproduced by using micropatterns composed of adhesive lines 12 μm wide connected to a circular disc of 50 μm in diameter. The total duration time is 5 min 50 s. The scale bar is 10 μm.

Supplementary Video 7

Time-lapse sequence in SiR-actin mode showing a single keratocyte that escaped from a narrow microstripe. The escape process was reproduced by using micropatterns composed of adhesive lines 12 μm wide connected to a circular disc of 50 μm in diameter. The total duration time is 4 min 25 s. The fluorescence intensity is colour-coded on 256 levels. The scale bar is 15 μm.

Supplementary Video 8

Time-lapse sequence in epifluorescent mode of a GFP–UTR zebrafish keratocyte (in green) migrating on a FN-coated microstripe 15 µm wide connected to a circular disc of 50 µm in diameter. The FN micropattern is labelled in red.

Supplementary Video 9

Time-lapse sequence in DIC mode of the migration of a train of cells (n = 2 cells) on a FN-coated microstripe 15 µm wide.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mohammed, D., Charras, G., Vercruysse, E. et al. Substrate area confinement is a key determinant of cell velocity in collective migration. Nat. Phys. 15, 858–866 (2019). https://doi.org/10.1038/s41567-019-0543-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41567-019-0543-3

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing