Project overview
This project aims to establish a biobank of airway epithelial cells from CF patients and matching control subjects. We are seeking funding to cover the consumable costs for specialised culture reagents and imaging techniques that are used to characterise the cell cultures. This project relies on human primary airway epithelial cells that will be isolated from airway brushings. During the duration of this project we aim to biobank airway epithelial cells from 25 volunteers (15 CF patients and 10 matching control subjects). We aim to recruit a higher number of CF patients as from our experience 1/3 of airway epithelial cell cultures fail to establish due to the increased bacterial load in these samples that cause a contamination and subsequently termination of cultures. Primary cells require highly defined culture media which are costly (£133.94/500ml). In order to characterise the differentiation of the cultures we are using immunofluorescence imaging techniques, requiring highly specialised reagents which are costly. We only request funds for antibodies specific for mucus producing cells (MUC5B: £281.50) and the tight junction protein ZO-1 (£246) as we currently don’t have these antibodies stocked in our lab. The usage of the confocal microscope will cost of approx. £20/h, we are accounting for 54h in total (4 full days plus 6h) for the duration of the project. Additionally, we will perform Transmission Electron Microscopy in order to analyse the structural organisation of the cell layer. As we are experienced in processing the samples ourselves we only request the time needed to analyse the samples with the electron microscope. We account for a total time of 4 hours at £67, which is a discounted rate offered by the Biomedical Imaging Unit. Overall, we estimate that the costs per donor account for £199 including media, plastic ware and characterisation of cultures using imaging.
