FIGURE 4.

Engraftment of decellularised dermis-based airway mucosal grafts in the chick chorioallantoic membrane assay (CAM). a) Timeline of the chick CAM assay. The egg is incubated and then windowed at Embryonic Day 3, while the scaffold is grafted at Embryonic Day 8 before harvest at Embryonic Day 10. b) Digital photographs of CAM assays after 2 days. c) Haematoxylin and eosin stained sections of the respiratory mucosal layer grafted onto CAM assay at Embryonic Day 10. An epithelial layer is demonstrated on the surface of the dermis overlying the CAM. Higher magnification of the epithelial layer demonstrates preservation of an epithelium but apparent loss of cilia. d) Immunofluorescence staining of sections of decellularised dermis-based respiratory mucosa. The 4′,6-diamidino-2-phenylindole (DAPI)-positive cell layer stained positively for the epithelial marker pan-keratin (green), the human cell marker STEM121 (yellow) and rarely the mucosecretory cell marker mucin 5AC (MUC5AC) (red). The CAM experiment involving epithelialised scaffolds was performed twice including six eggs in each experimental group on each occasion. The decellularised dermis CAM experiment was performed once with six eggs in each experimental group. All scale bars=50 µm. TE: tissue-engineered.