Substrates with Patterned Extracellular Matrix and Subcellular Stiffness Gradients Reveal Local Biomechanical Responses

Tseng, Peter; Carlo, Dino Di

doi:10.1002/adma.201304607

Cited by 44 publications

(38 citation statements)

References 40 publications

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“…Stress fibres are present along the four sides of the square envelope, and nonsupported fibres (NSF) are not characterized by a visible actin enrichment. Similar observations have been reported, showing that reinforced NSFs appear when the local stiffness increases 22 .…”

Section: Resultssupporting

confidence: 90%

Cell dipole behaviour revealed by ECM sub-cellular geometry

et al. 2014

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Cells sense and respond to their mechanical environment by exerting forces on their surroundings. The way forces are modulated by extra-cellular matrix (ECM) properties plays a key role in tissue homoeostasis. Using highly resolved micropatterns that constrain cells into the same square envelope but vary the adhesive geometry, here we investigate how the adhesive micro-environment affects the architecture of actin cytoskeleton and the orientation of traction forces. Our data demonstrate that local adhesive changes can trigger orientational ordering of stress fibres throughout the cell, suggesting that cells are capable of integrating information on ECM geometry at the whole-cell level. Finally, we show that cells tend to generate highly polarized force pattern, that is, unidirectional pinching, in response to adequate adhesive conditions. Hence, the geometry of adhesive environment can induce cellular orientation, a process which may have significant implications for the formation and mechanical properties of tissues.

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Section: Resultssupporting

confidence: 90%

Cell dipole behaviour revealed by ECM sub-cellular geometry

et al. 2014

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“…An important feature of this method is that it allows for independent selection of absolute high and low stiffness values and gradient steepness, accomplished by varying gel solution composition and gel dimensions, respectively. In previous studies where hydrogels with gradients in mechanical compliance were used to investigate durotaxis (31), the methods of generating mechanical gradients relied on juxtaposition of pregel solutions with varying concentrations of cross-linker (8,15,16), varying UV exposure in photopolymerization with a photomask (14, 19, 43), or overlaying a uniform gel solution on a rigid material with varying height to achieve a gradient in apparent rigidity at the surface (42,44). Though these methods effectively generate stiffness gradients, they are limited because they do not easily allow independent control of both the absolute stiffness range of the gels and the steepness of the generated gradient.…”

Section: Discussion Generation Of Hydrogels With Controlled Gradientsmentioning

confidence: 99%

Vascular smooth muscle cell durotaxis depends on extracellular matrix composition

Hartman

Isenberg

Chua

et al. 2016

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

121

110

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Mechanical compliance has been demonstrated to be a key determinant of cell behavior, directing processes such as spreading, migration, and differentiation. Durotaxis, directional migration from softer to more stiff regions of a substrate, has been observed for a variety of cell types. Recent stiffness mapping experiments have shown that local changes in tissue stiffness in disease are often accompanied by an altered ECM composition in vivo. However, the importance of ECM composition in durotaxis has not yet been explored. To address this question, we have developed and characterized a polyacrylamide hydrogel culture platform featuring highly tunable gradients in mechanical stiffness. This feature, together with the ability to control ECM composition, allows us to isolate the effects of mechanical and biological signals on cell migratory behavior. Using this system, we have tracked vascular smooth muscle cell migration in vitro and quantitatively analyzed differences in cell migration as a function of ECM composition. Our results show that vascular smooth muscle cells undergo durotaxis on mechanical gradients coated with fibronectin but not on those coated with laminin. These findings indicate that the composition of the adhesion ligand is a critical determinant of a cell's migratory response to mechanical gradients.durotaxis | cell migration | extracellular matrix | polyacrylamide C ell migration is essential to numerous biological processes, including development, angiogenesis, wound healing, and cancer metastasis (1-4). The movement of cells in these processes is determined by a complex assessment of environmental cues that include soluble factors, ECM composition, orientation, and stiffness. Numerous experiments have demonstrated that directional cell migration can result from gradients in these environmental cues: for example, chemotaxis (cell migration in response to gradients of soluble signals) and haptotaxis (cell migration in response to gradients of bound ligands) have been established in both in vitro and in vivo experimental systems (5-7). More recently, it has been demonstrated that cells are also capable of directed migration in response to gradients in substrate stiffness, a process termed durotaxis (8). Though there have been limited reports of specifically measured in vivo gradients (9), a number of recent stiffness mapping measurements imply the presence of stiffness gradients in both healthy and diseased tissues spanning a wide range of stiffnesses (10-13). In vitro experiments have demonstrated that directed migration in response to stiffness gradients can be observed in numerous cell types, using various materials as substrates, and across various stiffness levels (8,(14)(15)(16)(17)(18)(19). However, the role of ECM composition in mediating this behavior has not been thoroughly investigated.The interplay between mechanical stiffness and matrix composition in normal and pathological physiology is only now becoming appreciated. Recent studies in which tissue stiffness was mapped by atomic fo...

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“…This is of particular importance for studies focusing on complex, multivariate cell-ECM signaling pathways and the cytoskeletal response to different cell geometries and substrate stiffness 36 . Overall, local ECM density, cell shape, and substrate stiffness have been shown to regulate the structural organization of focal adhesion complexes 37,38 , the force balance between cell-cell and cell-ECM adhesions 4 , the nuclear lamina 39 , mesenchymal stem cell stiffness 40 , stem cell fate 12,41 , and the contractile properties of cardiomyocytes 42 .…”

Section: Discussionmentioning

confidence: 99%

Controlling cell shape on hydrogels using lift-off patterning

Moeller

Denisin

Sim

et al. 2017

Preprint

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Polyacrylamide gels functionalized with extracellular matrix proteins are commonly used as cell culture platforms to evaluate the combined effects of extracellular matrix composition, cell geometry and substrate rigidity on cell physiology. For this purpose, protein transfer onto the surface of polyacrylamide hydrogels must result in geometrically well-resolved micropatterns with homogeneous protein distribution. Yet the outcomes of micropatterning methods have not been pairwise evaluated against these criteria. We report a high-fidelity photoresist lift-off patterning method to pattern ECM proteins on polyacrylamide hydrogels ranging from 5 to 25 kPa. We directly compare the protein transfer efficiency and pattern geometrical accuracy of this protocol to the widely used microcontact printing method. Lift-off patterning achieves higher protein transfer efficiency, increases pattern accuracy, increases pattern yield, and reduces variability of these factors within arrays of patterns as it bypasses the drying and transfer steps of microcontact printing. We demonstrate that lift-off patterned hydrogels successfully control cell size and shape and enable long-term imaging of actin intracellular structure and lamellipodia dynamics when we culture epithelial cells on these substrates.

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Substrates with Patterned Extracellular Matrix and Subcellular Stiffness Gradients Reveal Local Biomechanical Responses

Cited by 44 publications

References 40 publications

Cell dipole behaviour revealed by ECM sub-cellular geometry

Cell dipole behaviour revealed by ECM sub-cellular geometry

Vascular smooth muscle cell durotaxis depends on extracellular matrix composition

Controlling cell shape on hydrogels using lift-off patterning

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