Do Amniotic Fluid Cells Represent a Viable Cell Source for Regeneration of the Neonatal Upper Airway? (360G-Wellcome-201265_Z_16_Z)

Aim To evaluate human Amniotic Fluid Cells (AFCs) for fetal and neonatal tracheobronchial tissue engineering. Objectives & Methodology I will isolate hAFC subpopulations already displaying primitive airway basal cell surface markers and quantify co-expression of nuclear respiratory lineage markers (NKx2.1, FOXA2 and SOX2). I will ascertain if this subpopulation can be enriched using ‘proximal airway’ culture conditions, and whether these enriched hAFCs can be rapidly expanded via our novel in-house epithelial expansion protocol. I will investigate whether my enriched ‘respiratory’ hAFC population can differentiate to form cell types present in mature tracheobronchial epithelium by manipulation of Wnt, Notch and TGF-beta signalling. I will assess their ability to generate a ciliated pseudostratified epithelium in vitro in ALI and 3D spheroid cultures. I will use an immunosuppressed rabbit airway surgical model to assess the in vivo suitability of these airway-differentiated hAFCs. I will implant seeded decellularised rabbit donor tracheae in lateral thoracic musculofascial flaps and, following orthotopic transfer of grafts, I will analyse the grafts weekly (by bronchoscopic biopsy) and at post-mortem using histology and immunofluorescence. Keywords Tissue Engineering - Regenerative Medicine – Fetal and Neonatal Therapies - Amniotic Fluid Cells – Autologous Cell Source - Proximal Airway Epithelium - Epithelial Expansion