Fig. 5. Sorting keratinocytes for a ß1^bri/Dsg3^dim population further enhanced the enrichment of colony forming cells. (A) Keratinocytes dual labelled for ß1 integrin and Dsg3 were sorted into two groups: ß1^bri/Dsg3^dim (11.5% of the cells, with the highest ß1 and least Dsg3) and ß1^bri/Dsg3^bri (5.39% of the cells, with both the highest ß1 and Dsg3) populations (a). (b,c) FSC and SSC plot of ß1^bri/Dsg3^dim and ß1^bri/Dsg3^bri populations, respectively. Note that 85% of cells in ß1^bri/Dsg3^dim (b) but only 40% of cells in ß1^bri/Dsg3^bri (c) have low SSC characteristics. Relatively more cells in c show high FSC and SSC characteristics. (B) Sorted sole (thick skin; a) and breast (thin skin; b) keratinocytes (Kc) were plated onto the feeders at 1000 or 500 cells per well, respectively, and grown for 11 days. Colonies were stained for keratins and visualised using anti-mouse HRP. There are significantly more colonies in ß1^bri/Dsg3^dim than ß1^bri/Dsg3^bri wells of both cell types. (C) Quantitative colony analyses include colony density (total colony area per dish) and CFE (ratio of colony number to plated cell number) (left graph), colony size (mean colony area) and colony number (mean colony number per dish) (right graph). There is a 4- or 5-fold increase in colony density, and a 7- or 3-fold increase in CFE in ß1^bri/Dsg3^dim compared with ß1^bri/Dsg3^bri in either thick or thin skin keratinocytes, respectively (^*P<0.05). Significantly more colonies are seen in ß1^bri/Dsg3^dim rather than ß1^bri/Dsg3^bri population (^*P<0.05) and larger colonies show in thin skin Kc (^*P<0.05).