Traction force microscopy – Measuring the forces exerted by cells

Lekka, Małgorzata; Gnanachandran, Kajangi; Kubiak, Andrzej; Zieliński, Tomasz; Zemła, Joanna

doi:10.1016/j.micron.2021.103138

Cited by 64 publications

(44 citation statements)

References 101 publications

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“…The current gold standard for assessing isolated cell actomyosin activity is traction force microscopy (TFM) ( Muhamed et al, 2017 ). TFM is used to quantitatively measure the stress exerted by a cell on its substrate, which is then used as an indicator of cell contractility ( Kraning-Rush et al, 2012 ; Lekka et al, 2021 ). Additionally, atomic force microscopy (AFM) has been used to assess actomyosin cortical tension and the force transduced through individual focal adhesion complexes ( Sanyour et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Using Polyacrylamide Hydrogels to Model Physiological Aortic Stiffness Reveals that Microtubules Are Critical Regulators of Isolated Smooth Muscle Cell Morphology and Contractility

et al. 2022

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Vascular smooth muscle cells (VSMCs) are the predominant cell type in the medial layer of the aortic wall and normally exist in a quiescent, contractile phenotype where actomyosin-derived contractile forces maintain vascular tone. However, VSMCs are not terminally differentiated and can dedifferentiate into a proliferative, synthetic phenotype. Actomyosin force generation is essential for the function of both phenotypes. Whilst much is already known about the mechanisms of VSMC actomyosin force generation, existing assays are either low throughput and time consuming, or qualitative and inconsistent. In this study, we use polyacrylamide hydrogels, tuned to mimic the physiological stiffness of the aortic wall, in a VSMC contractility assay. Isolated VSMC area decreases following stimulation with the contractile agonists angiotensin II or carbachol. Importantly, the angiotensin II induced reduction in cell area correlated with increased traction stress generation. Inhibition of actomyosin activity using blebbistatin or Y-27632 prevented angiotensin II mediated changes in VSMC morphology, suggesting that changes in VSMC morphology and actomyosin activity are core components of the contractile response. Furthermore, we show that microtubule stability is an essential regulator of isolated VSMC contractility. Treatment with either colchicine or paclitaxel uncoupled the morphological and/or traction stress responses of angiotensin II stimulated VSMCs. Our findings support the tensegrity model of cellular mechanics and we demonstrate that microtubules act to balance actomyosin-derived traction stress generation and regulate the morphological responses of VSMCs.

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Section: Introductionmentioning

confidence: 99%

Using Polyacrylamide Hydrogels to Model Physiological Aortic Stiffness Reveals that Microtubules Are Critical Regulators of Isolated Smooth Muscle Cell Morphology and Contractility

et al. 2022

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“…Moreover, measurements at this length scale are different from tissue-level mechanical properties. Macroscopic characterization methods such as traction force microscopy, rheometry, and elastic micropillars have been used to measure forces at supracellular length scale [9][10][11] .…”

Section: Introductionmentioning

confidence: 99%

Magneto-acoustic protein nanostructures for non-invasive imaging of tissue mechanics in vivo

Kim

Min

Kim

et al. 2022

Preprint

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Measuring cellular and tissue mechanics inside intact living organisms is essential for interrogating the roles of force in physiological and disease processes, and is a major goal in the field of mechanobiology. However, existing biosensors for 3D tissue mechanics, primarily based on fluorescent emissions and deformable materials, are limited for in vivo measurement due to the limited light penetration and poor material stability inside intact, living organisms. While magneto-motive ultrasound (MMUS), which uses superparamagnetic nanoparticles as imaging contrast agents, has emerged as a promising modality for real-time in vivo imaging of tissue mechanics, it has poor sensitivity and spatiotemporal resolution. To overcome these limitations, we introduce magneto-gas vesicles (MGVs), a unique class of protein nanostructures based on gas vesicles and magnetic nanoparticles that produces differential ultrasound signals in response to varying mechanical properties of surrounding tissues. These hybrid protein nanostructures significantly improve signal strength and detection sensitivity. Furthermore, MGVs enable non-invasive, long-term, and quantitative measurement of mechanical properties within 3D tissues and organs in vivo. We demonstrated the performance of MGV-based mechano-sensors in vitro, in fibrosis models of organoids, and in vivo in mouse liver fibrosis models.

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“…Here we use well-characterized and reproducible ultra-soft polyacrylamide gels (PAGs) of variable elasticity (0.4-200kPa) to quantify the stresses exerted by T cells during their early spreading (first 15-30 minutes). We employ Traction force microscopy (TFM, (Style et al 2014; Lekka et al 2021)) to follow the dynamics, magnitude, and directionality of the stresses generated during the first minutes of T cell engagement with PAGs of stiffness similar to that of non-activated DCs and B cells, that is to say, about 400 Pa (Bufi et al 2015). The cells are specifically engaged via the CD3 domain of the TCR complex, and/or the co-receptor CD28, and/or the T cell integrin LFA.…”

Section: Introductionmentioning

confidence: 99%

T Cell Traction Forces show temporal patterns which Depend on Stimulation and Cell Subtype

Mustapha

Pelicot-Biarnes

Torro

et al. 2022

Preprint

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T cells use forces to read out and act on the mechanical parameters of their microenvironment, which includes antigen presenting cells (APCs). Here we explore the early interaction of T cells with an APC-mimicking ultra-soft polymer gel exhibiting physiologically relevant stiffness in the range of 350-450 Pa. We quantify the dependence of cell spreading and stiffness on gel elasticity, and measure early time traction forces. We find that coating the surface with an antibody against the CD3 region of the TCR-complex elicits small but measurable gel deformation in the early recognition phase, which we quantify in terms of stress or energy. We show that the time evolution of the energy follows one of three distinct patterns: active fluctuation, intermittent, or sigmoidal signal. Addition of either anti-CD28 or anti-LFA1 has little impact on the total integrated energy or the maximum stress. However, the relative distribution of the energy patterns does depend on the additional ligands. Remarkably, the forces are centrifugal at very early times, and only later turn into classical in-ward pointing centripetal traction.

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Traction force microscopy – Measuring the forces exerted by cells

Cited by 64 publications

References 101 publications

Using Polyacrylamide Hydrogels to Model Physiological Aortic Stiffness Reveals that Microtubules Are Critical Regulators of Isolated Smooth Muscle Cell Morphology and Contractility

Using Polyacrylamide Hydrogels to Model Physiological Aortic Stiffness Reveals that Microtubules Are Critical Regulators of Isolated Smooth Muscle Cell Morphology and Contractility

Magneto-acoustic protein nanostructures for non-invasive imaging of tissue mechanics in vivo

T Cell Traction Forces show temporal patterns which Depend on Stimulation and Cell Subtype

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