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• W3201065629 abstract "The topic of this thesis are cell mechanical responses to large forces. This topic is relevant because many cell types in the human body are permanently subjected to large deformations or mechanical forces. However, current methods to investigate mechanical properties at the cellular level are mostly limited to small forces and deformations, and to cell behavior in the linear regime. This thesis explores the magnetic tweezer microrheometer as a method to extend measurements to the non-linear regime of cell mechanical behavior up to the level of cell or adhesion bond breakage. In the course of experiments with an improved high-force magnetic tweezer device on living cells, several discoveries were made, including the discovery that the structural plasticity of cells is coupled to the elastic and dissipative cell properties measured in the linear regime, and that the adhesion complex of living cells shows catch bond behavior. To understand the molecular and structural origin of those behaviors, the magnetic tweezer method was combined with other state-of-the-art methods for measuring cell forces and deformations.This dissertation is divided into two main parts. In the first part, the magnetic tweezer, the cell stretcher, and the 2D traction force cytometry method are introduced. Moreover, method improvements and the implementation of device extensions are described. This includes the expansion of the force range and the application of multi-directional forces with the magnetic tweezer, a new protocol for 2D traction force microscopy measurements in order to achieve better reproducibility and higher throughput, as well as an essential improvement of cell stretcher experiments to analyze the fraction of cells that lost adhesion with the substrate, as opposed to cells that have undergone cell death.The second part describes the application of these methods and presents results of cell experimental studies on cell plasticity and cell adhesion dynamics. A central result is the discovery of a constitutive equation of cell mechanical behavior in the high-force regime that accurately predicts the mechanical response of cells to large, multiple and long-lasting forces, including the response after the forces have been removed. Moreover, a novel force spectroscopy method was implemented with the high-force magnetic tweezer: when forces acting on magnetic beads that are coupled to cell adhesion complexes are large, the adhesion bond can break. In the course of the experiments it became evident that the dynamics of bond breakage did not follow the classical Bell-type bond dynamics but instead it followed a catch bond behavior. Such catch bond behavior has been previously shown for single protein-protein adhesion bonds but not for the adhesion complex of living cells.To exclude that the observed bond-breakage dynamics was caused by a complex bond energy landscape, a novel staircase-like force protocol was introduced that confirmed catch bond behavior as the dominant source of adhesion bond breakage dynamics. In addition, the data were compared with Monte-Carlo simulations that supported the finding of a catch bond behavior. To investigate the molecular basis of cell plasticity and catch bond-type focal adhesions, experiments with cell stretcher, 2D traction force microscopy as well as a high-force magnetic tweezer were carried out in cells carrying mutations of proteins of the cytoskeleton and the focal adhesion complex, including NEDD9, filamin A, desmin, and plectin. Results show that these proteins caused pronounced mechanical alterations in cells, but none were solely responsible for the plastic cell responses or the catch bond behavior of cell adhesions, in line with the interpretation that both properties are robust, highly redundant and fundamental features of living cell." @default.
• W3201065629 created "2021-09-27" @default.
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• W3201065629 date "2014-01-01" @default.
• W3201065629 modified "2023-09-24" @default.
• W3201065629 title "Cell Mechanics in Response to Large Forces and Deformations" @default.
• W3201065629 hasPublicationYear "2014" @default.
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