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2018 ; 114
(11
): 2743-2755
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Collective Cell Behavior in Mechanosensing of Substrate Thickness
#MMPMID29874622
Tusan CG
; Man YH
; Zarkoob H
; Johnston DA
; Andriotis OG
; Thurner PJ
; Yang S
; Sander EA
; Gentleman E
; Sengers BG
; Evans ND
Biophys J
2018[Jun]; 114
(11
): 2743-2755
PMID29874622
show ga
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 ?m) 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 ?m. 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 ?m. 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.