Dynamic traction force measurements of migrating immune cells in 3D matrices

Böhringer, David; Cóndor, Mar; Bischof, Lars; Czerwinski, Tina; Bauer, Andreas; Voskens, Caroline J.; Budday, Silvia; Mark, Christoph; Fabry, Ben; Gerum, Richard

doi:10.1101/2022.11.16.516758

Cited by 6 publications

(16 citation statements)

References 60 publications

(71 reference statements)

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“…To test how the contractility of the NK92 cells changes in response to a varying matrix stiffness, we reconstruct the cell contractility from the measured deformation fields. Importantly, we employ a material model that specifically includes non-affine deformations and accurately captures the non-linear effects of strain stiffening and buckling of collagen fibers 8,14 . As immune cells are expected to migrate without substantial force generation most of the time, we focus on the maximum contractility that we obtain within the 24 min measurement period of each cell.…”

Section: Contractility and Mechanosensitivity Of Nk92 Cellsmentioning

confidence: 99%

“…Batch results in stiffer collagen gels (storage modulus 286 Pa) and is used for Figures 2, 4, 7a-b, 7g-h and Supplementary Figure 7, while batch 2 (shear modulus 84 Pa) is used for Figures 3, 5, 6, 7c-d and Supplementary Figure 4. Rheological measurements are conducted as previously described in Böhringer et al 14 In brief, collagens hydrogels (85µl) are polymerized for 30 minutes at 37°C between the plates of a shear rheometer (Discovery HR-3, TA Instruments, Milford, 20mm cone-plate geometry). Subsequently, a frequency sweep (0.2-2Hz) is performed at a low amplitude of 1%, and the stress-strain relationship is measured (0-100% strain) with a constant strain rate of 1%/s.…”

Section: Collagen Gel Preparationmentioning

confidence: 99%

“…Pore diameters for both batches is 4.4 µm as determined from confocal reflection image stacks (volume of 160x160x200 µm with a voxel sizes of 0.314x0.314x0.642 µm) as described in Refs. 14,35,36…”

Section: Collagen Gel Preparationmentioning

confidence: 99%

“…3D traction force microscopy (Fig. 2b-e, 3, 4a-c, 5a,b, 5d-f, 6) is conducted as described in Böhringer et al 14 and Condor et al 38 . In brief, we pipette 3 ml collagen (0.6 -2.4 mg/ml) solution with 200,000 cells into a 35 mm Nunc cell culture treated Petri dish (ThermoFisher Scientific).…”

Section: Single Cell 3-d Traction Force Microscopymentioning

confidence: 99%

“…We select elongated cells for traction force measurements specifically, as these are assumed to be motile. From the recorded 3D reflection-channel image stacks, we extract the cell-induced deformations of the collagen fibers using 3D particle image velocimetry as implemented by the SAENOPY Python software package 14 . Based on these 3D deformation fields, SAENOPY computes the contractility of each cell at each measured time step.…”

Section: Single Cell 3-d Traction Force Microscopymentioning

confidence: 99%

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Immune cells employ intermittent integrin-mediated traction forces for 3D migration

Czerwinski

Bischof

Böhringer

et al. 2023

Preprint

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To reach targets outside the bloodstream, immune cells can extravasate and migrate through connective tissue. However, in contrast to migrating mesenchymal cells, the importance of matrix adhesion and traction force generation for immune cell migration is not well understood. We use time-lapse confocal reflection microscopy to obtain simultaneous measurements of migration velocity, directional persistence, and cell contractility. While we confirm that immune cells use a non-contractile amoeboid migration mode by default, we also find that NK92 cells as well as ex-vivo expanded NK cells exert substantial acto-myosin driven contractile forces on the extracellular matrix during short contractile phases reaching up to 100nN. Even non-activated primary B, T, NK cells, neutrophils, and monocytes exhibit this burst-like contractile behavior, and NK activation with cytokines increases both the magnitude and frequency of contractile bursts. Importantly, we show that cell speed and directional persistence of NK cells increase during and after these contractile phases, implying that the cells actively use traction forces to overcome steric hindrance and avoid getting stuck in narrow pores of the ECM. Accordingly, reducing cell adhesion to the ECM reduces the fraction of motile cells and their directional persistence, while the remaining motile cells mostly maintain their cell speed. We conclude that steric hindrance can induce a switch in the migration mode of immune cells, from a non-adhesive amoeboid migration mode to a highly contractile migration mode that closely resembles the gliding motion of motile mesenchymal cells.

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Section: Contractility and Mechanosensitivity Of Nk92 Cellsmentioning

confidence: 99%

Section: Collagen Gel Preparationmentioning

confidence: 99%

Section: Collagen Gel Preparationmentioning

confidence: 99%

Section: Single Cell 3-d Traction Force Microscopymentioning

confidence: 99%

Section: Single Cell 3-d Traction Force Microscopymentioning

confidence: 99%

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Immune cells employ intermittent integrin-mediated traction forces for 3D migration

Czerwinski

Bischof

Böhringer

et al. 2023

Preprint

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Facile Determination of the Poisson’s Ratio and Young’s Modulus of Polyacrylamide Gels and Polydimethylsiloxane

Smith,

Inocencio,

Pardi

et al. 2024

ACS Appl. Polym. Mater.

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Polyacrylamide hydrogels (PAH gel) and polydimethylsiloxane (PDMS, an elastomer) are two soft materials often used in cell mechanics and mechanobiology, in manufacturing lab-on-a-chip applications, among others. This is partly due to the ability to tune their elasticity with ease in addition to various chemical modifications. For affine polymeric networks, two (of three) elastic constants, Young's modulus (E), the shear modulus (G), and Poisson's ratio (ν), describe the purely elastic response to external forces. However, the literature addressing the experimental determination of ν for PAH (sometimes called PAA gels in the literature) and the PDMS elastomer is surprisingly limited when compared to the literature that reports values of the elastic moduli, E and G. Here, we present a facile method to obtain the Poisson's ratio and Young's modulus for PAH gel and PDMS elastomer based on static tensile tests. The value of ν obtained from the deformation of the sample is compared to the value determined by comparing E and G via a second independent method that utilizes small amplitude shear rheology. We show that the Poisson's ratio may vary significantly from the value for incompressible materials (ν = 0.5), often assumed in the literature even for soft compressible hydrogels. Surprisingly, we find a high degree of agreement between elastic constants obtained by shear rheology and macroscopic static tension test data for polyacrylamide hydrogels but not for elastomeric PDMS.

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T cells use focal adhesions to pull themselves through confined environments

Caillier,

Oleksyn,

Fowell

et al. 2024

Journal of Cell Biology

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Immune cells are highly dynamic and able to migrate through environments with diverse biochemical and mechanical compositions. Their migration has classically been defined as amoeboid under the assumption that it is integrin independent. Here, we show that activated primary Th1 T cells require both confinement and extracellular matrix proteins to migrate efficiently. This migration is mediated through small and dynamic focal adhesions that are composed of the same proteins associated with canonical mesenchymal cell focal adhesions, such as integrins, talin, and vinculin. These focal adhesions, furthermore, localize to sites of contractile traction stresses, enabling T cells to pull themselves through confined spaces. Finally, we show that Th1 T cells preferentially follow tracks of other T cells, suggesting that these adhesions modify the extracellular matrix to provide additional environmental guidance cues. These results demonstrate not only that the boundaries between amoeboid and mesenchymal migration modes are ambiguous, but that integrin-mediated focal adhesions play a key role in T cell motility.

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Dynamic traction force measurements of migrating immune cells in 3D matrices

Cited by 6 publications

References 60 publications

Immune cells employ intermittent integrin-mediated traction forces for 3D migration

Immune cells employ intermittent integrin-mediated traction forces for 3D migration

Facile Determination of the Poisson’s Ratio and Young’s Modulus of Polyacrylamide Gels and Polydimethylsiloxane

T cells use focal adhesions to pull themselves through confined environments

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