Multi-oscillation microrheology <i>via</i> acoustic force spectroscopy enables frequency-dependent measurements on endothelial cells at high-throughput

Nguyen, Alfred; Brandt, Matthias; Muenker, Till M.; Betz, Timo

doi:10.1039/d0lc01135e

Cited by 26 publications

(37 citation statements)

References 53 publications

(41 reference statements)

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“…Human umbilical vein endothelial cells (HUVECs) were a kind gift from Prof. Dr. V. Gerke (Institute of Medical Biochemistry, University of Münster, Germany). The cells were cultured in 60 mm diameter polystyrene dishes (Corning CellBIND Surface 60 mm Culture Dish, Corning) in culture medium consisting of two equal parts of Endothelial Cell Growth Medium 2 (PromoCell) containing its according supplements (Growth Medium 2 SupplementMix, PromoCell) and of Medium 199 Earle's (F0615, Biochrom GmbH or M2154, Sigma-Aldrich) with 2.2 g/l NaHCO 3 , without L-glutamine supplemented with 10 % fetal bovine serum (F7524, Sigma-Aldrich), 0.2 units/ml heparin (H3149, Sigma), 30 µg/ml gentamicin (gibco) and 15 µg/ml amphotericin B (gibco) as previously described [46]. Culture dishes were placed in a humidified incubator at 37 • and 5 % CO 2 .…”

Section: Cell Culture and Treatmentsmentioning

confidence: 99%

Human endothelial cells display a rapid and fluid flow dependent tensional stress increase in response to tumor necrosis factor-α

Brandt¹,

Gerke²,

Betz

2022

Preprint

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As endothelial cells form the inner layer of blood vessels they display the first barrier to interstitial tissues, which results in a crucial role for inflammation. On the global, systemic level an important element of the complex process controlling the inflammatory response is the release of the cytokine tumor necrosis factor-α (TNF-α). While other pro-inflammatory agents like thrombin or histamine are known to induce acute but transient changes in endothelial cells which have been well studied biologically as well as mechanically, TNF-α is primarily known for its sustained effects on permeability and leukocyte recruitment. These functions are associated with transcriptional changes that take place on the timescale of hours and days. Here we show that already 4 minutes after the addition of TNF-α onto monolayers of human umbilical vein endothelial cells, a striking rise in mechanical substrate traction force and internal monolayer tension can be recorded. As expected, the traction forces act primarily at the boundary of the monolayer. While the traction forces increase monotonically during the initial cellular response, we find that the internal monolayer tension displays a rapid peak that can be abolished when applying a shear flow to the cells. The increased internal monolayer tension may provide a mechanical signal for the cells to prepare for the recruitment of leukocytes, additionally to the well studied biochemical response.

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Section: Cell Culture and Treatmentsmentioning

confidence: 99%

Human endothelial cells display a rapid and fluid flow dependent tensional stress increase in response to tumor necrosis factor-α

Brandt¹,

Gerke²,

Betz

2022

Preprint

Self Cite

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show abstract

“…Oscillatory microrheology fixes the timescale of the perturbation according to the applied frequency, while the other tests probe a superposition on many timescales in a single test. This has the advantage that the frequency-dependency of the material response can be measured directly and that the measurement frequency can be adopted to the frequency range of interest [52]. However, timeand frequency-domain data are mathematical equivalents and time-domain data of the cells viscoelastic response can even be directly transformed into the frequency domain [53].…”

Section: Discussionmentioning

confidence: 99%

Oscillatory Microrheology, Creep Compliance and Stress Relaxation of Biological Cells Reveal Strong Correlations as Probed by Atomic Force Microscopy

et al. 2021

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The mechanical properties of cells are important for many biological processes, including wound healing, cancers, and embryogenesis. Currently, our understanding of cell mechanical properties remains incomplete. Different techniques have been used to probe different aspects of the mechanical properties of cells, among them microplate rheology, optical tweezers, micropipette aspiration, and magnetic twisting cytometry. These techniques have given rise to different theoretical descriptions, reaching from simple Kelvin-Voigt or Maxwell models to fractional such as power law models, and their combinations. Atomic force microscopy (AFM) is a flexible technique that enables global and local probing of adherent cells. Here, using an AFM, we indented single retinal pigmented epithelium cells adhering to the bottom of a culture dish. The indentation was performed at two locations: above the nucleus, and towards the periphery of the cell. We applied creep compliance, stress relaxation, and oscillatory rheological tests to wild type and drug modified cells. Considering known fractional and semi-fractional descriptions, we found the extracted parameters to correlate. Moreover, the Young’s modulus as obtained from the initial indentation strongly correlated with all of the parameters from the applied power-law descriptions. Our study shows that the results from different rheological tests are directly comparable. This can be used in the future, for example, to reduce the number of measurements in planned experiments. Apparently, under these experimental conditions, the cells possess a limited number of degrees of freedom as their rheological properties change.

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“…This is essential as Nguyen et al. extensively showed that the force distribution in the chip can vary depending on the position of the bead ( Nguyen et al., 2021 ). The spatial calibration map (n = 20 beads) for our field of view obtained by performing the Stokes force calibration is shown in Figure S2 .…”

Section: Methodsmentioning

confidence: 99%

Single-cell analysis reveals chemokine-mediated differential regulation of monocyte mechanics

et al. 2022

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Summary Monocytes continuously adapt their shapes for proper circulation and elicitation of effective immune responses. Although these functions depend on the cell mechanical properties, the mechanical behavior of monocytes is still poorly understood and accurate physiologically relevant data on basic mechanical properties are lacking almost entirely. By combining several complementary single-cell force spectroscopy techniques, we report that the mechanical properties of human monocyte are strain-rate dependent, and that chemokines can induce alterations in viscoelastic behavior. In addition, our findings indicate that human monocytes are heterogeneous mechanically and this heterogeneity is regulated by chemokine CCL2. The technology presented here can be readily used to reveal mechanical complexity of the blood cell population in disease conditions, where viscoelastic properties may serve as physical biomarkers for disease progression and response to therapy.

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Multi-oscillation microrheology via acoustic force spectroscopy enables frequency-dependent measurements on endothelial cells at high-throughput

Abstract: Active microrheology is one of the main methods to determine the mechanical properties of cells and tissue, and the modelling of these viscoelastic properties is under heavy debate with many...

Cited by 26 publications

References 53 publications

Human endothelial cells display a rapid and fluid flow dependent tensional stress increase in response to tumor necrosis factor-α

Human endothelial cells display a rapid and fluid flow dependent tensional stress increase in response to tumor necrosis factor-α

Oscillatory Microrheology, Creep Compliance and Stress Relaxation of Biological Cells Reveal Strong Correlations as Probed by Atomic Force Microscopy

Single-cell analysis reveals chemokine-mediated differential regulation of monocyte mechanics

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