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Collective cell behavior in mechanosensing of substrate thickness

Collective cell behavior in mechanosensing of substrate thickness
Collective cell behavior in mechanosensing of substrate thickness

Extracellular matrix stiffness has a profound effect on the behavior of many cell types. Adherent cells apply contractile forces to the material on which they adhere and sense the resistance of the material to deformation—its stiffness. This is dependent on both the elastic modulus and the thickness of the material, with the corollary that single cells are able to sense underlying stiff materials through soft hydrogel materials at low (<10 μm) thicknesses. Here, we hypothesized that cohesive colonies of cells exert more force and create more hydrogel deformation than single cells, therefore enabling them to mechanosense more deeply into underlying materials than single cells. To test this, we modulated the thickness of soft (1 kPa) elastic extracellular-matrix-functionalized polyacrylamide hydrogels adhered to glass substrates and allowed colonies of MG63 cells to form on their surfaces. Cell morphology and deformations of fluorescent fiducial-marker-labeled hydrogels were quantified by time-lapse fluorescence microscopy imaging. Single-cell spreading increased with respect to decreasing hydrogel thickness, with data fitting to an exponential model with half-maximal response at a thickness of 3.2 μm. By quantifying cell area within colonies of defined area, we similarly found that colony-cell spreading increased with decreasing hydrogel thickness but with a greater half-maximal response at 54 μm. Depth-sensing was dependent on Rho-associated protein kinase-mediated cellular contractility. Surface hydrogel deformations were significantly greater on thick hydrogels compared to thin hydrogels. In addition, deformations extended greater distances from the periphery of colonies on thick hydrogels compared to thin hydrogels. Our data suggest that by acting collectively, cells mechanosense rigid materials beneath elastic hydrogels at greater depths than individual cells. This raises the possibility that the collective action of cells in colonies or sheets may allow cells to sense structures of differing material properties at comparatively large distances.

0006-3495
2743-2755
Tusan, Camelia G.
729621e2-3f9f-4c0a-9800-99e0afeaf673
Man, Yu Hin
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Zarkoob, Hoda
4c2febf0-4b96-417e-b31f-ea492955d51c
Johnston, David A.
b41163c9-b9d2-425c-af99-2a357204014e
Andriotis, Orestis G.
d88047ee-bc23-4b1b-9d98-184685fefb65
Thurner, Philipp J.
ab711ddd-784e-48de-aaad-f56aec40f84f
Yang, Shoufeng
e0018adf-8123-4a54-b8dd-306c10ca48f1
Sander, Edward A.
1da883da-29d0-46e2-8386-834f322070c5
Gentleman, Eileen
a0b4959e-7b65-46b7-9ea8-1d0f5857c22f
Sengers, Bram G.
d6b771b1-4ede-48c5-9644-fa86503941aa
Evans, Nicholas D.
06a05c97-bfed-4abb-9244-34ec9f4b4b95
Tusan, Camelia G.
729621e2-3f9f-4c0a-9800-99e0afeaf673
Man, Yu Hin
e4534f4d-3531-4483-9f70-562b8bffd987
Zarkoob, Hoda
4c2febf0-4b96-417e-b31f-ea492955d51c
Johnston, David A.
b41163c9-b9d2-425c-af99-2a357204014e
Andriotis, Orestis G.
d88047ee-bc23-4b1b-9d98-184685fefb65
Thurner, Philipp J.
ab711ddd-784e-48de-aaad-f56aec40f84f
Yang, Shoufeng
e0018adf-8123-4a54-b8dd-306c10ca48f1
Sander, Edward A.
1da883da-29d0-46e2-8386-834f322070c5
Gentleman, Eileen
a0b4959e-7b65-46b7-9ea8-1d0f5857c22f
Sengers, Bram G.
d6b771b1-4ede-48c5-9644-fa86503941aa
Evans, Nicholas D.
06a05c97-bfed-4abb-9244-34ec9f4b4b95

Tusan, Camelia G., Man, Yu Hin, Zarkoob, Hoda, Johnston, David A., Andriotis, Orestis G., Thurner, Philipp J., Yang, Shoufeng, Sander, Edward A., Gentleman, Eileen, Sengers, Bram G. and Evans, Nicholas D. (2018) Collective cell behavior in mechanosensing of substrate thickness. Biophysical Journal, 114 (11), 2743-2755. (doi:10.1016/j.bpj.2018.03.037).

Record type: Article

Abstract

Extracellular matrix stiffness has a profound effect on the behavior of many cell types. Adherent cells apply contractile forces to the material on which they adhere and sense the resistance of the material to deformation—its stiffness. This is dependent on both the elastic modulus and the thickness of the material, with the corollary that single cells are able to sense underlying stiff materials through soft hydrogel materials at low (<10 μm) thicknesses. Here, we hypothesized that cohesive colonies of cells exert more force and create more hydrogel deformation than single cells, therefore enabling them to mechanosense more deeply into underlying materials than single cells. To test this, we modulated the thickness of soft (1 kPa) elastic extracellular-matrix-functionalized polyacrylamide hydrogels adhered to glass substrates and allowed colonies of MG63 cells to form on their surfaces. Cell morphology and deformations of fluorescent fiducial-marker-labeled hydrogels were quantified by time-lapse fluorescence microscopy imaging. Single-cell spreading increased with respect to decreasing hydrogel thickness, with data fitting to an exponential model with half-maximal response at a thickness of 3.2 μm. By quantifying cell area within colonies of defined area, we similarly found that colony-cell spreading increased with decreasing hydrogel thickness but with a greater half-maximal response at 54 μm. Depth-sensing was dependent on Rho-associated protein kinase-mediated cellular contractility. Surface hydrogel deformations were significantly greater on thick hydrogels compared to thin hydrogels. In addition, deformations extended greater distances from the periphery of colonies on thick hydrogels compared to thin hydrogels. Our data suggest that by acting collectively, cells mechanosense rigid materials beneath elastic hydrogels at greater depths than individual cells. This raises the possibility that the collective action of cells in colonies or sheets may allow cells to sense structures of differing material properties at comparatively large distances.

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Accepted/In Press date: 20 March 2018
e-pub ahead of print date: 7 June 2018

Identifiers

Local EPrints ID: 421860
URI: http://eprints.soton.ac.uk/id/eprint/421860
ISSN: 0006-3495
PURE UUID: a38ffbb6-670b-4fee-84ef-1f855125c693
ORCID for David A. Johnston: ORCID iD orcid.org/0000-0001-6703-6014
ORCID for Philipp J. Thurner: ORCID iD orcid.org/0000-0001-7588-9041
ORCID for Shoufeng Yang: ORCID iD orcid.org/0000-0002-3888-3211
ORCID for Bram G. Sengers: ORCID iD orcid.org/0000-0001-5859-6984

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Date deposited: 03 Jul 2018 16:30
Last modified: 18 Feb 2021 17:15

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Contributors

Author: Camelia G. Tusan
Author: Yu Hin Man
Author: Hoda Zarkoob
Author: David A. Johnston ORCID iD
Author: Orestis G. Andriotis
Author: Shoufeng Yang ORCID iD
Author: Edward A. Sander
Author: Eileen Gentleman
Author: Bram G. Sengers ORCID iD

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