Three-dimensional force microscopy of cells in biopolymer networks

Steinwachs, Julian; Metzner, Claus; Skodzek, Kai; Lang, Nadine; Thievessen, Ingo; Mark, Christoph; Münster, Stefan; Aifantis, Katerina E.; Fabry, Ben

doi:10.1038/nmeth.3685

Cited by 288 publications

(372 citation statements)

References 39 publications

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“…Our results show that MDA-MB-231 cells exhibiting mesenchymal motility displace their surrounding collagen matrix much more extensively than previously reported for a mixed population of amoeboid and mesenchymal cells (Fig. S2B) (42). We note that fibroblasts exhibit even stronger strain-stiffening behavior than cancer cells of epithelial origin as reported in refs.…”

Section: From Cell To Ecm: Cells Exert Sufficient Strain To Locally Asupporting

confidence: 83%

Fibrous nonlinear elasticity enables positive mechanical feedback between cells and ECMs

Hall

Alisafaei

Ban

et al. 2016

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

297

282

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In native states, animal cells of many types are supported by a fibrous network that forms the main structural component of the ECM. Mechanical interactions between cells and the 3D ECM critically regulate cell function, including growth and migration. However, the physical mechanism that governs the cell interaction with fibrous 3D ECM is still not known. In this article, we present single-cell traction force measurements using breast tumor cells embedded within 3D collagen matrices. We recreate the breast tumor mechanical environment by controlling the microstructure and density of type I collagen matrices. Our results reveal a positive mechanical feedback loop: cells pulling on collagen locally align and stiffen the matrix, and stiffer matrices, in return, promote greater cell force generation and a stiffer cell body. Furthermore, cell force transmission distance increases with the degree of strain-induced fiber alignment and stiffening of the collagen matrices. These findings highlight the importance of the nonlinear elasticity of fibrous matrices in regulating cell-ECM interactions within a 3D context, and the cell force regulation principle that we uncover may contribute to the rapid mechanical tissue stiffening occurring in many diseases, including cancer and fibrosis.cell traction force | 3D cell traction force microscopy | fibrous nonlinear elasticity | cell-ECM interaction | collagen A nimal cells of most cell types, including breast tumor cells, are supported structurally by a fibrous ECM within a 3D context (1, 2). Cells adhere to the fibers via the linkages between integrin receptors on the membrane surface and the adhesion molecules within the ECM. To migrate, cells pull/push along the fibers or squeeze through the pore structure of the network (3). The tensional balance between the cell and the ECM critically regulates many physiological and pathological processes, including immune response, tissue formation, and tumor progression (4-7). In the breast tumor, stiffening of the mechanical environment disrupts force balance between epithelial cells and the ECM, promoting a malignant phenotype (5, 8). Tumors stiffen as cells deposit more collagen than they digest (9-11), increasingly express cross-linking enzymes (8, 12), and exert traction forces to reorganize the ECM (13).The main structural component of the ECM is a network of cross-linked protein fibers. The fiber network aligns, stiffens, and sometimes, undergoes permanent changes when subjected to strain (14, 15). These adaptive mechanical properties of the fiber network provide cells entry points to modify their local microenvironment (16-18) and as such, perform physiologically realistic functions (1,3,(19)(20)(21)(22)(23). It has been reported that the nonlinear elasticity of fibrous matrices enables cells to transmit forces over distances of hundreds of micrometers, facilitating long-range communication between individual cells (24-26) and between tumor spheroids (27). Recent work has shown that individual cells are capable of stiffening the...

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Section: From Cell To Ecm: Cells Exert Sufficient Strain To Locally Asupporting

confidence: 83%

Fibrous nonlinear elasticity enables positive mechanical feedback between cells and ECMs

Hall

Alisafaei

Ban

et al. 2016

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

297

282

View full text Add to dashboard Cite

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“…The tangent elastic modulus between 3 and 7% axial strains (∼0.16 to ∼0.75 kPa) at 0.5, 1.0, and 2.0 mg/mL collagen concentrations is also shown from ref. 53. (D) Quantitative analysis of the melanoma cell invasion shows a biphasic response to the concentration of collagen.…”

Section: The Interplay Between Cell Contractility and Matrix Fiber Rementioning

confidence: 99%

Modeling the two-way feedback between contractility and matrix realignment reveals a nonlinear mode of cancer cell invasion

Ahmadzadeh

Webster

Behera

et al. 2017

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

164

151

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Cancer cell invasion from primary tumors is mediated by a complex interplay between cellular adhesions, actomyosin-driven contractility, and the physical characteristics of the extracellular matrix (ECM). Here, we incorporate a mechanochemical free-energy-based approach to elucidate how the two-way feedback loop between cell contractility (induced by the activity of chemomechanical interactions such as Ca 2+ and Rho signaling pathways) and matrix fiber realignment and strain stiffening enables the cells to polarize and develop contractile forces to break free from the tumor spheroids and invade into the ECM. Interestingly, through this computational model, we are able to identify a critical stiffness that is required by the matrix to break intercellular adhesions and initiate cell invasion. Also, by considering the kinetics of the cell movement, our model predicts a biphasic invasiveness with respect to the stiffness of the matrix. These predictions are validated by analyzing the invasion of melanoma cells in collagen matrices of varying concentration. Our model also predicts a positive correlation between the elongated morphology of the invading cells and the alignment of fibers in the matrix, suggesting that cell polarization is directly proportional to the stiffness and alignment of the matrix. In contrast, cells in nonfibrous matrices are found to be rounded and not polarized, underscoring the key role played by the nonlinear mechanics of fibrous matrices. Importantly, our model shows that mechanical principles mediated by the contractility of the cells and the nonlinearity of the ECM behavior play a crucial role in determining the phenotype of the cell invasion.cell invasion | cell contractility | matrix realignment | Rho pathway | fibrous matrices C ell invasion into the surrounding matrix from nonvascularized primary tumors is the main mechanism by which cancer cells migrate to nearby blood vessels and metastasize to eventually form secondary tumors. This process is mediated by an intricate intercoupling between intracellular forces (such as cell contractility) and extracellular forces (adhesions and protrusions) that depend on the stiffness of the surrounding stroma and the alignment of matrix fibers. Previous experimental studies have examined the influence of these forces on the migratory behavior of cells during invasion. For example, the comparison between cell contractility in malignant and normal tissues has shown that the cells with malignant phenotype have a higher level of contractility (1-4). This elevated contractility is directly proportional to factors such as the stiffness of the extracellular matrix (ECM) and the fiber realignment (5-7), suggesting that the cross talk between ECM and intracellular contractility mediated by mechanosensory signaling pathways is also implicated in metastasis. Specifically, the activity of Rho, a myosin GTPase that regulates the activity of myosins, is elevated in proportion to the stiffness of the surrounding matrix (1,8,9), and inhibition of Rho-associated ...

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“…More recently Steinwachs and colleagues have developed a physio-mimetic biopolymer based system that allows the direct quantification of the mechanical forces applied by cancer cells fully embedded within physiological relevant 3D network [63]. Another recent study has reported the use of stimulated emission depletion (STED) microscopy with TFM to solve one of the major shortcomings of this technique, the spatial resolution.…”

Section: Traction Force Microscopymentioning

confidence: 99%

Pancreatic cancer: a mechanobiology approach

Chronopoulos

Lieberthal

Hernández

2017

Converg. Sci. Phys. Oncol.

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Despite many years of effort from cancer biologists and clinical oncologists, pancreatic ductal adenocarcinoma (PDAC) remains a thoroughly recalcitrant disease, resisting both conventional forms of cancer treatment and radical innovative therapies. Perhaps driving the lack of success in PDAC is the underappreciation of the primary hallmark of the tumour: a fibrotic extracellular matrix (ECM) that composes a majority of the highly rigid solid carcinoma. In recent years we have come to understand that the homeostasis of ECM mechanics is pivotal for the homeostasis of cells and the progression or initiation of a malignant phenotype. This can be understood by way of mechanobiology, a field that attempts to understand how physical forces like ECM stiffness alter cell behaviour. In this review, we provide an overview of our understanding of PDAC from this perspective and the recent advances in biophysics and engineering that allow for new tools with which to investigate the mechanobiology of PDAC.

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Three-dimensional force microscopy of cells in biopolymer networks

Cited by 288 publications

References 39 publications

Fibrous nonlinear elasticity enables positive mechanical feedback between cells and ECMs

Fibrous nonlinear elasticity enables positive mechanical feedback between cells and ECMs

Modeling the two-way feedback between contractility and matrix realignment reveals a nonlinear mode of cancer cell invasion

Pancreatic cancer: a mechanobiology approach

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