Traction microscopy to identify force modulation in subresolution adhesions

Han, Sangyoon J.; Oak, Youbean; Groisman, Alex; Danuser, Gaudenz

doi:10.1038/nmeth.3430

Cited by 155 publications

(226 citation statements)

References 34 publications

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“…4B). Such a value corresponds to the range of stress that cells are able to generate on their substrate 41,42 and therefore supports the relevance of the present system in mimicking the mechanical coupling of neighbouring cells through their matrix 43 . In terms of force, magneto-active substrates can locally transmit nN-forces to cell adhesions, if a typical adhesion surface of 1µm 2 is considered, which also compares to forces generated by single adhesions 44,45 .…”

Section: Generation Of the Magnetic Fieldsupporting

confidence: 78%

Magneto-Active Substrates for Local Mechanical Stimulation of Living Cells

Coullomb

Bidan

Fratzl

et al. 2018

Biophysical Journal

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Cells are able to sense and react to their physical environment by translating a mechanical cue into an intracellular biochemical signal that triggers biological and mechanical responses. This process, called mechanotransduction, controls essential cellular functions such as proliferation and migration. The cellular response to an external mechanical stimulation has been investigated with various static and dynamic systems, so far limited to global deformations or to local stimulation through discrete substrates. To apply local and dynamic mechanical constraints at the single cell scale through a continuous surface, we have developed and modelled magneto-active substrates made of magnetic micro-pillars embedded in an elastomer. Constrained and unconstrained substrates are analysed to map surface stress resulting from the magnetic actuation of the micro-pillars and the adherent cells. These substrates have a rigidity in the range of cell matrices, and the magnetic micro-pillars generate local forces in the range of cellular forces, both in traction and compression. As an application, we followed the protrusive activity of cells subjected to dynamic stimulations. Our magneto-active substrates thus represent a new tool to study mechanotransduction in single cells, and complement existing techniques by exerting a local and dynamic stimulation, traction and compression, through a continuous soft substrate.Living cells have a sense of touch, which means that they are able to feel, respond and adapt to the mechanical properties of their environment. The process by which cells convert mechanical signals into biochemical signals is called mechanotransduction. Defects in the mechanotransduction pathways are implicated in numerous diseases ranging from atherosclerosis and osteoporosis to cancer progression and developmental disorders 1,2 . Since the 1990s, different static studies focused on mechanosensing have shown that cells can migrate along the rigidity gradient direction 3 and that stem cells can differentiate in vitro according to their substrate's stiffness 4 and geometry 5 . The interplay between a mechanical force and the reinforcement of cell adhesion has also been documented 6,7 . In their natural environment, cells face a complex and dynamic mechanical environment. Cyclic strain can induce reorientation of adherent cells and affect cell growth depending on the temporal and spatial properties of the mechanical stimulation [8][9][10][11] . The relevant timescales span from the milli-second for the stretching of mechanosensitive proteins, minutes for mechanotransduction signalling to hours for global morphological changes and even longer for adapting cell functions 12 . Taken together, previous works have shown that cells are sensitive to both the spatial and temporal signatures of mechanical stimuli. In order to study mechanotransduction, it is thus essential to stimulate cells with mechanical cues controlled both spatially and temporally.To address this topic, various methods have been proposed to exert experim...

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Section: Generation Of the Magnetic Fieldsupporting

confidence: 78%

Magneto-Active Substrates for Local Mechanical Stimulation of Living Cells

Coullomb

Bidan

Fratzl

et al. 2018

Biophysical Journal

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“…For this reason, intense efforts have been devoted to understanding how key signaling molecules and ECM characteristics influence the formation and growth of FAs. In particular, in vitro studies using elastic hydrogels have shown that forces generated by actomyosin contraction are essential for the stabilization of FAs (5,6). Numerous observations have convincingly demonstrated that cells form larger FAs as well as develop higher intracellular traction forces on stiffer ECMs (7,8), evidencing the mechanosensitive nature of FAs which has been quantitatively modeled using different (continuum, coarse-grain, and molecular) approaches (9,10).…”

mentioning

confidence: 99%

Multiscale model predicts increasing focal adhesion size with decreasing stiffness in fibrous matrices

Cao

Ban

Baker

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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We describe a multiscale model that incorporates force-dependent mechanical plasticity induced by interfiber cross-link breakage and stiffness-dependent cellular contractility to predict focal adhesion (FA) growth and mechanosensing in fibrous extracellular matrices (ECMs). The model predicts that FA size depends on both the stiffness of ECM and the density of ligands available to form adhesions. Although these two quantities are independent in commonly used hydrogels, contractile cells break cross-links in soft fibrous matrices leading to recruitment of fibers, which increases the ligand density in the vicinity of cells. Consequently, although the size of focal adhesions increases with ECM stiffness in nonfibrous and elastic hydrogels, plasticity of fibrous networks leads to a departure from the well-described positive correlation between stiffness and FA size. We predict a phase diagram that describes nonmonotonic behavior of FA in the space spanned by ECM stiffness and recruitment index, which describes the ability of cells to break cross-links and recruit fibers. The predicted decrease in FA size with increasing ECM stiffness is in excellent agreement with recent observations of cell spreading on electrospun fiber networks with tunable cross-link strengths and mechanics. Our model provides a framework to analyze cell mechanosensing in nonlinear and inelastic ECMs.focal adhesion | mechanosensing | cell contractility | matrix physical remodeling | Rho pathway F ocal adhesions (FAs) are large macromolecular assemblies through which mechanical force and regulatory signals are transmitted between the extracellular matrix (ECM) and cells. FAs play important roles in many cellular behaviors, including proliferation, differentiation, and locomotion, and pathological processes like tumorigenesis and wound healing (1-4). For this reason, intense efforts have been devoted to understanding how key signaling molecules and ECM characteristics influence the formation and growth of FAs. In particular, in vitro studies using elastic hydrogels have shown that forces generated by actomyosin contraction are essential for the stabilization of FAs (5, 6). Numerous observations have convincingly demonstrated that cells form larger FAs as well as develop higher intracellular traction forces on stiffer ECMs (7,8), evidencing the mechanosensitive nature of FAs which has been quantitatively modeled using different (continuum, coarse-grain, and molecular) approaches (9, 10).It must be pointed out that in all of the aforementioned investigations, the substrates considered were flat (2D) and linear elastic. However, in vivo, many cells reside within 3D fibrous scaffolds where the density and diameter of fibers can vary depending on the nature of the tissue (11-13). The local architecture of these fibrous networks may change significantly when cells exert forces on them, leading to phenomena such as nonlinear stiffening, reorientation, and physical remodeling of the ECM (14, 15). Our recent study on cells in synthetic fibrous matrices wit...

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“…Common substrates used for TFM include polyacrylamide or silicon based gels. They are selected due to their linear elasticity, optical transparency, and tunability of elastic moduli through polymer chain crosslinking, over several orders of magnitude [95]. Knowing the elastic moduli of the substrates, the displacement field can be converted to a traction force field using an inverse method [93, 96,97].…”

Section: Techniques To Study Cell–cell Adhesionmentioning

confidence: 99%

Techniques to stimulate and interrogate cell–cell adhesion mechanics

Yang

Broussard

Green

et al. 2018

Extreme Mechanics Letters

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Cell–cell adhesions maintain the mechanical integrity of multicellular tissues and have recently been found to act as mechanotransducers, translating mechanical cues into biochemical signals. Mechanotransduction studies have primarily focused on focal adhesions, sites of cell-substrate attachment. These studies leverage technical advances in devices and systems interfacing with living cells through cell–extracellular matrix adhesions. As reports of aberrant signal transduction originating from mutations in cell–cell adhesion molecules are being increasingly associated with disease states, growing attention is being paid to this intercellular signaling hub. Along with this renewed focus, new requirements arise for the interrogation and stimulation of cell–cell adhesive junctions. This review covers established experimental techniques for stimulation and interrogation of cell–cell adhesion from cell pairs to monolayers.

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Traction microscopy to identify force modulation in subresolution adhesions

Cited by 155 publications

References 34 publications

Magneto-Active Substrates for Local Mechanical Stimulation of Living Cells

Magneto-Active Substrates for Local Mechanical Stimulation of Living Cells

Multiscale model predicts increasing focal adhesion size with decreasing stiffness in fibrous matrices

Techniques to stimulate and interrogate cell–cell adhesion mechanics

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