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Characterisation of cultured airway basal cells to understand their role in human lung disease

Hynds, RE; (2016) Characterisation of cultured airway basal cells to understand their role in human lung disease. Doctoral thesis , UCL (University College London). Green open access

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Abstract

Many studies in murine models have demonstrated the stem/progenitor cell potential of basal epithelial cells in the tracheal epithelium. However, significant differences exist between the respiratory epithelium in rodents and in man. As such, novel methodologies to study respiratory epithelial cells in vitro are in demand. Here, methods to expand primary human airway epithelial cells from living patients were explored. The field’s ‘gold standard’ medium for the expansion of these cells was poorly suited to initiating cultures from small endobronchial biopsy samples as proliferation of these cells was time-limited and after a short period of time in culture the cells became senescent and were unable to regenerate a mucociliary epithelium in organotypic models. As such, an alternative epithelial culture strategy involving the co-culture of human airway epithelial cells with 3T3-J2 fibroblast feeder cells in medium containing a small molecule Rho-associated protein kinase (ROCK) inhibitor was assessed. This method greatly improved both the yield and the longevity of human basal cell cultures and allowed multipotent airway differentiation in organotypic assays after longer culture periods than conventional techniques. Finally, the epithelial-stromal cell crosstalk between epithelial cells and feeder cells in co-culture was investigated, revealing a novel signalling pathway involving phosphorylation of the transcription factor signal transducer and activator of transcription 6 (STAT6) by hepatocyte growth factor (HGF) signalling.

Type: Thesis (Doctoral)
Title: Characterisation of cultured airway basal cells to understand their role in human lung disease
Event: University College London
Open access status: An open access version is available from UCL Discovery
Language: English
Keywords: Lung, stem cells, epithelium, cell culture
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute > Research Department of Oncology
URI: http://discovery.ucl.ac.uk/id/eprint/1503706
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