Traction Force Microscopy in 3‐Dimensional Extracellular Matrix Networks

Cóndor, Mar; Steinwachs, Julian; Mark, Christoph; García-Aznar, J.M.; Fabry, Ben

doi:10.1002/cpcb.24

Cited by 32 publications

(31 citation statements)

References 13 publications

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“…For a final collagen concentration of 1.2 mg/ml, we dilute the solution before polymerization with a mixture of one volume part NaHCO 3 , one part 10× cRPMI, and eight parts H 2 O (ref. 30 ) and adjust the solution to pH 9 with NaOH. After polymerization at 37 °C, 5% CO 2 , and 95% RH for 1 h, 1.5 ml of RPMI medium (for primary NK cells) or 1.5 ml of Alpha-MEM medium (for NK92 cells) is added to each well of a six-well plate.…”

Section: Methodsmentioning

confidence: 99%

Cryopreservation impairs 3-D migration and cytotoxicity of natural killer cells

et al. 2020

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Natural killer (NK) cells are important effector cells in the immune response to cancer. Clinical trials on adoptively transferred NK cells in patients with solid tumors, however, have thus far been unsuccessful. As NK cells need to pass stringent safety evaluation tests before clinical use, the cells are cryopreserved to bridge the necessary evaluation time. Standard degranulation and chromium release cytotoxicity assays confirm the ability of cryopreserved NK cells to kill target cells. Here, we report that tumor cells embedded in a 3-dimensional collagen gel, however, are killed by cryopreserved NK cells at a 5.6-fold lower rate compared to fresh NK cells. This difference is mainly caused by a 6-fold decrease in the fraction of motile NK cells after cryopreservation. These findings may explain the persistent failure of NK cell therapy in patients with solid tumors and highlight the crucial role of a 3-D environment for testing NK cell function.

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Section: Methodsmentioning

confidence: 99%

Cryopreservation impairs 3-D migration and cytotoxicity of natural killer cells

et al. 2020

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“…More precisely, passive cell mechanical analyses are performed in the absence of well-defined local forces and analyze the fluctuations of particles or structural elements or the deformation of materials by cells. Among these techniques are the nanoscale particle tracking (Bursac et al, 2005; Mierke, 2011a), membrane fluctuation measurements or flicker spectroscopy (bending stiffness) (Döbereiner et al, 2003; Loftus et al, 2013), traction force measurements on PAA gels (Mierke et al, 2008a, b, 2011a,b) or micropillars (Heil and Spatz, 2010; Schoen et al, 2010) and matrix displacement analysis (Fischer et al, 2017; Kunschmann et al, 2019) or matrix bead displacement analysis (Franck et al, 2011; Steinwachs et al, 2016; Cóndor et al, 2017), when cells themselves migrate and invade through a 3D confined extracellular matrix (Figure 2). The limitations associated with mechanical techniques pose challenges to the detection of high-resolution spatiotemporal alterations of cells or cell compartments.…”

Section: Cell Mechanical Probing Techniques and Adhesive State Of A Cellmentioning

confidence: 99%

The Role of the Optical Stretcher Is Crucial in the Investigation of Cell Mechanics Regulating Cell Adhesion and Motility

Mierke

2019

Front. Cell Dev. Biol.

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The mechanical properties of cells, tissues, and the surrounding extracellular matrix environment play important roles in the process of cell adhesion and migration. In physiological and pathological processes of the cells, such as wound healing and cancer, the capacity to migrate through the extracellular matrix is crucial. Hence biophysical techniques were used to determine the mechanical properties of cells that facilitate the various migratory capacities. Since the field of mechanobiology is rapidly growing, the reliable and reproducible characterization of cell mechanics is required that facilitates the adhesion and migration of cells. One of these cell mechanical techniques is the optical stretching device, which was originally developed to investigate the mechanical properties of cells, such as the deformation of single cells in suspension. After discussing the strengths and weaknesses of the technology, the latest findings in optical stretching-based cell mechanics are presented in this review. Finally, the mechanical properties of cells are correlated with their migratory potential and it is pointed out how the inhibition of biomolecules that contribute to the to the maintenance of cytoskeletal structures in cells affect their mechanical deformability.

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“…In this second case, collagen gels were not fluorescently labelled and were analysed by means of confocal reflection microscopy. These two methods are currently the most used for the study of collagen microstructures in 3D in biological laboratories (Chung et al, 2012;Cóndor et al, 2017;Del Amo et al, 2018;Kueng et al, 1989;Leclerc et al, 2003;Sung et al, 2009).…”

Section: Preparation Of Collagen Gelsmentioning

confidence: 99%

“…After polymerization, 2 ml of complete cell culture medium was added to prevent dehydration of collagen gels. For more information about the critical parameters and troubleshooting for generating collagen gels, see (Cóndor et al, 2017). These gels were used for the characterization of matrix structure around cells.…”

Section: Collagen Gels Plated In Cell Culture Dishesmentioning

confidence: 99%

Image-based Characterization of 3D Collagen Networks and the Effect of Embedded Cells

et al. 2019

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Collagen microstructure is closely related to the mechanical properties of tissues and affects cell migration through the extracellular matrix. To study these structures, three-dimensional (3D) in vitro collagen-based gels are often used, attempting to mimic the natural environment of cells. Some key parameters of the microstructure of these gels are fiber orientation, fiber length, or pore size, which define the mechanical properties of the network and therefore condition cell behavior. In the present study, an automated tool to reconstruct 3D collagen networks is used to extract the aforementioned parameters of gels of different collagen concentration and determine how their microstructure is affected by the presence of cells. Two different experiments are presented to test the functionality of the method: first, collagen gels are embedded within a microfluidic device and collagen fibers are imaged by using confocal fluorescence microscopy; second, collagen gels are directly polymerized in a cell culture dish and collagen fibers are imaged by confocal reflection microscopy. Finally, we investigate and compare the collagen microstructure far from and in the vicinities of MDA-MB 23 cells, finding that cell activity during migration was able to strongly modify the orientation of the collagen fibers and the porosity-related values.

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Traction Force Microscopy in 3‐Dimensional Extracellular Matrix Networks

Cited by 32 publications

References 13 publications

Cryopreservation impairs 3-D migration and cytotoxicity of natural killer cells

Cryopreservation impairs 3-D migration and cytotoxicity of natural killer cells

The Role of the Optical Stretcher Is Crucial in the Investigation of Cell Mechanics Regulating Cell Adhesion and Motility

Image-based Characterization of 3D Collagen Networks and the Effect of Embedded Cells

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