Fig. 2

Characterisation of NFCs in vivo and after initial FDGS treatment. Two-day-old neonatal mouse (A) and two-week-old Adult mouse (B) dorsal skin immunolabelled with K14 antibody (red). Inserts show higher magnification views of the epidermal component. C Confocal micrographs of the neonatal mouse skin whole-mount immunolabelled with K14 (red) and Sox2 (green). Inserts show higher magnification views of DP. All sections with DAPI nuclear counterstain (blue). Scale bars: 100 μm. D H&E stain of dorsal skin showing HF morphogenesis stage and DP sphere formation. Stage 3: Dermal fibroblasts form a rounded DP cell. Stage 4: DP cells are greater in length than width and are enclosed by > 50%. Stage 5: DP cells are almost completely encapsulated and ball-shaped. Asterisks denote DP-spheres. E The schematic diagram of FDGS 1st procedure. F Photographs of a bilayer discontinuous Ficoll density gradient overlayed with dermal mixed cells in medium before and after centrifugation. G Representative images of NFC, before and after FDGS1st treatment, show a significant reduction in non-NFC single cells. Scale bars: 50 μm