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Measurement of mechanical tractions exerted by cells in three-dimensional matrices

Nature Methods volume 7, pages 969–971 (2010)Cite this article

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Abstract

Quantitative measurements of cell-generated forces have heretofore required that cells be cultured on two-dimensional substrates. We describe a technique to quantitatively measure three-dimensional traction forces exerted by cells fully encapsulated in well-defined elastic hydrogel matrices. Using this approach we measured traction forces for several cell types in various contexts and revealed patterns of force generation attributable to morphologically distinct regions of cells as they extend into the surrounding matrix.

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Figure 1: Cell-induced hydrogel deformations and construction of a discretized Green's function.
Figure 2: Measurement of tractions exerted by live cells.
Figure 3: Measurement of dynamic tractions exerted by spreading cells.

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Acknowledgements

We thank J. Baranski, H. Hu, C. Shen and M. Wozniak for helpful discussions. This work was supported in part by grants from the US National Institutes of Health (EB00262, EB08396, GM74048, HL73305 and HL90747), the Resbio Technology Resource for Polymeric Biomaterials, the Material Research Science and Engineering Center and Center for Engineering Cells and Regeneration at the University of Pennsylvania, the National Science Foundation graduate research fellowship (W.R.L. and B.L.B.), the US National Institutes of Health T32 training grant and the Hartwell Foundation (J.S.M.).

Author information

Authors and Affiliations

  1. Department of Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania, USA

    Wesley R Legant, Jordan S Miller, Brandon L Blakely, Daniel M Cohen & Christopher S Chen

  2. Department of Mechanical, Aerospace and Structural Engineering, Washington University in St. Louis, St. Louis, Missouri, USA

    Guy M Genin

Authors
  1. Wesley R Legant
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  2. Jordan S Miller
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  3. Brandon L Blakely
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  4. Daniel M Cohen
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Contributions

W.R.L., G.M.G. and C.S.C. conceived and initiated the project. W.R.L., J.S.M., B.L.B. and D.M.C. designed and performed experiments. C.S.C. supervised the project.

Corresponding author

Correspondence to Christopher S Chen.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–10, Supplementary Notes 1–3 (PDF 3523 kb)

Supplementary Movie 1

Volume rendering of encapsulated cell and surrounding beads. An EGFP-expressing fibroblast (green) was encapsulated within a PEGDA hydrogel containing fluorescent beads (red). After spreading into the surrounding hydrogel, the cell was imaged using confocal microscopy. For clarity, the cell was rendered in front of the beads and only one half of all beads were rendered. (MOV 9907 kb)

Supplementary Movie 2

A 3D wall-eyed stereogram of bead displacements. Volume rendering showing the discretized surface mesh of the cell and tracked bead displacements, color-coded by magnitude, obtained from bead locations before and after cell lysis. (MOV 9641 kb)

Supplementary Movie 3

Cell lysis and hydrogel relaxation. A 2D confocal section showing an EGFP-expressing fibroblast and surrounding fluorescent beads during cell lysis with SDS. (MOV 4272 kb)

Supplementary Movie 4

A 3D wall-eyed stereogram rendering of cellular tractions. Tractions are color-coded by magnitude. For clarity, the surface mesh was made transparent to permit visualization of traction vectors directed inward from the cell surface. (MOV 8125 kb)

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Legant, W., Miller, J., Blakely, B. et al. Measurement of mechanical tractions exerted by cells in three-dimensional matrices. Nat Methods 7, 969–971 (2010). https://doi.org/10.1038/nmeth.1531

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