Abstract:
Cells are the basic structural and functional units of all living things (Tortora, 2006). Cells constantly interact with their surrounding environment. One such form of interaction is a cell’s ability to adhere or anchor itself to its surroundings. Adherent cells can generate what are known as traction forces on its surroundings through protein structures known as focal adhesions (DuFort, 2011). Cells in vivo are subjected to a range of forces and can adjust its focal adhesions, cytoskeletal organisation, overall state and also its traction forces to respond to these changes (Discher, 2005). One such example, is how endothelial cells sense and respond to shear stress generated from the frictional force of flowing blood (Ando, 2013). The key mechanisms by which these cells respond to shear forces, have yet to be identified (Ando, 2013). The aim of this project was to develop a validated, traction force microscopy flow system for the accurate measurement of cellular traction forces exerted by endothelial cells under flow. This system will allow future investigation into the nature of endothelial cell to environment interactions and the cellular and molecular mechanisms behind many basic biological processes such as cell migration and adhesion at both the cell and tissue levels (Plotnikov, 2014). During this project, a traction force microscopy flow system was set up, where key components of the system were validated. The validation experiments carried out showed promising results in: the even distribution of extracellular matrix protein on the substrate; the tracking of particle displacement; and the verification of the effect of low shear flow on the surface of the substrate. However, before this system can be used in research to measure the traction forces exerted by endothelial cells under the effect of shear flow, an accurate method of gel characterization must be incorporated into this system and the flow system has to be validated for high shear stresses.
