Figure 4
Analysis of engraftment and proliferation of CB-CD34-derived cells. A-C: Alu-FISH analysis. A: Human postmortem lung tissue (positive control) showing nuclear positivity in all cells. B: Representative micrograph of lungs of animal treated with CD34+ cells expanded in basic medium showing a cluster of alu-FISH-positive cells within airspace. C. Lung of animal treated with DEX-expanded cells showing collection of alu-FISH-positive cells along the alveolar wall, admixed with larger-sized, FISH-negative murine alveolar macrophages. A-C: alu-FISH analysis (green) with DAPI counterstain (blue). D-F: Alu-FISH analysis combined with anti-Ki67 immunofluorescence. D. Representative micrograph of animal treated with freshly isolated CD34+ cells, showing relatively large-sized proliferating alu-FISH positive cells within the alveolar wall (arrows). E. Lungs of animal treated with CD34+ cells expanded in basic medium showing a large cluster of alu-FISH-positive cells free within the airspace. Proliferative activity is present in these CD34-derived cells (arrow). F. Lungs of animal treated with DEX-expanded CD34+ cells, similarly showing Ki67-positivity in FISH-positive intraalveolar cells (arrow). Several proliferating FISH-negative murine cells are noted. D-F: alu-FISH analysis (green) combined with anti-Ki67 immunofluorescence (red); DAPI counterstain (blue).