Development of bioengineered vascular model for functional vascular genomics

About the Project

Second Supervisor: Dr Tom Webb, Department of Cardiovascular Sciences, University of Leicester

‘Omics’ technologies such as transcriptomics, proteomics and epigenomics analyse biological molecules on a large scale, providing comprehensive insights into the structure, function, and dynamics of organisms. When applied on cultured cells these technologies enable insight into fundamental mechanisms of health and disease. There is a significant limitation though; cells are generally grown in monolayers on plastic, an environment which bares little relation to the in vivo environment. Our interest is particularly the vascular environment, here cells are organised in three-dimensional structures, interact with neighbouring cell types and the surrounding extra-cellular matrix, and are exposed to dynamic mechanical forces, particularly shear stress from blood flow. This project turns to biomedical engineering to create an optimised 3d culture environment; key structural proteins will be combined with multiple vascular cell types in a mimic of the native vessel environment. Biomaterials will provide structural and chemical cues for cell arrangement and support physiologically relevant shear and pulsatile forces in a single scalable platform with:

• 3D vessel-like architecture
• Co-culture capability for multiple vascular cell types
• Unidirectional and customisable flow to mimic physiological and pathological shear stress
• Sufficient cell yield for comprehensive omic profiling
• Compatibility with live imaging platforms for phenotyping

By creating such a model, we aim to create a transformative platform for functional vascular genomics - one that bridges the gap between genetic discoveries and mechanistic insights, paving the way for new targeted therapeutic strategies. More broadly such a representative culture environment supports the goal of 3Rs (refinement, reduction replacement,) in animal studies for both mechanistic understanding and in drug development.

Techniques that will be undertaken during the project

• Creation of multi-layer vascular constructs
• A range of a materials processing techniques including, lyophilisation, film casting, electrospinning and fibre extrusion
• Advanced characterisation of constructs prior to culture – electron microscopy, raman microscopy, x-ray micro-computed tomography, mechanical testing and simulation of physiological shear loading.
• Cell seeding and co-culture methodologies within multi-layer vascular construct
• Focus on seeding optimisation – advanced 3D imaging and IF
• Combination of construct with pulsatile flow reactor
• Vascular cell culture (standard 2D static culture and 3D culture)
• Biomolecule extraction from cultured cells (DNA, RNA and protein, as applicable)
• Microscopy and related techniques (e.g. immunofluorescence) for monitoring 3D cell culture and phenotypic effects
• ChIP-seq library preparation and data analysis ·
• ATAC-seq library preparation and data analysis

Enquiries

Project Enquiries to js1005@leicester.ac.uk

To apply please use the application link at the bottom of this web page Engineering | Postgraduate research | University of Leicester

Funding Notes

This project is only available on a self funded basis or if you have your own sponsorship.