Figure 2

Flow cytometry of limbal cells and sorting. (A) Flow cytometry analyses of cluster derived limbal cells showing a high percentage of CD117⁺/Melan-A⁺ cells compared to stromal fraction-derived cells. Stromal fraction-derived cells also showing the presence of CD117⁺/Melan-A⁻ cells. Percentages (%) of positive cells are expressed as the means ± SEM (n = 4). (B) Fluorescent cell sorting of cluster derived cells (i, P0–0 day), enrichment of CD117⁺ cells showing a gradual increase (i, P0–10 days), finally yielded a pure population of CD117⁺ cells (i, P1–10 days). Percentages (%) of positive cells are expressed as the means ± SEM percentage (%) (n = 7) (i). Phase-contrast images showing the attached multidendritic cells after 24 h of seeding of CD117⁺ cells (ii, P0–1 day) and stromal contamination after a few days in culture (black arrows) (ii, P0–10 days). Sorting of cultured cells showing the absence of fibroblast and all cells showing large bodies, flattened, smooth, multiple dendritic morphologies (ii, P1–10 days). Magnification × 100.