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Development of a novel gene screening platform for cells exposed to flow

Moers, B (2017) Development of a novel gene screening platform for cells exposed to flow. MPhil thesis, University of Sheffield.

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Abstract

Flow is a key regulator of endothelial function and therefore cardiovascular disease development. Nonetheless, the application of flow to cultured cells is a non-trivial challenge. Results obtained from in vitro flow systems must be critically reviewed as several technical limitations exist within each system. Recent developments of molecular high throughput techniques are constantly changing the scientific demands of life science laboratories. In order to meet this demand, there should also be the possibility to increase the throughput for flow studies. However, at present there is no high throughput device commercially available to apply flow to cells in vitro. Therefore there is a strong need for a novel high throughput technology that can effectively monitor the impact of shear stress on cells. Endothelial cells (EC) form the inner lining of the arterial wall, thus acting as a barrier between the vessel wall and flowing blood. EC are subjected to wall shear stress (WSS), a force created by the flowing blood. Although Atherosclerosis is associated with systematic risk factors like Diabetes or smoking, disease characteristic plaques occur predominantly near branches and bends, where EC are exposed to low or bidirectional WSS; thus illustrating the physiological importance of WSS. This project aims to develop a high throughput in vitro system for functional screening of gene regulation in EC exposed to flow. The development of an in vitro device for application of flow to cells in 96 well plates will allow screening to identify genes involved in cellular responses to flow. It is envisaged that the system will be used in future studies to identify novel putative therapeutic targets for the prevention or treatment of atherosclerosis. Therefore, the proposed high throughput device is designed to be compatible with standard cell culture methods. Briefly, flow will be generated in a standard 96 well cell culture plate using 96 individual stirring cones mounted in a re-usable lid. Each cone will be rotated by a magnetic stirrer, placed underneath the 96 well plate. Due to the device’s compatibility with plate readers it will be an ideal addition to existing flow studies to perform fluorescent in situ staining.

Item Type: Thesis (MPhil)
Academic Units: The University of Sheffield > Faculty of Engineering (Sheffield)
The University of Sheffield > Faculty of Engineering (Sheffield) > Mechanical Engineering (Sheffield)
Depositing User: MS B Moers
Date Deposited: 12 Feb 2018 09:38
Last Modified: 12 Feb 2018 09:38
URI: http://etheses.whiterose.ac.uk/id/eprint/19386

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