A: Morphological analysis using HPS staining, Ki67, K15, and filaggrin expression. K15 is rarely expressed when the skin is reconstructed using fibroblasts alone, weakly with 50%/50% density and strongly with a density of 75% fibroblasts and 25% ASCs. Filaggrin, was highly expressed when 25% ASCs were added to the fibroblasts during skin reconstruction and weakly expressed when 50% were added. On the other hand, in control epidermis with 100% fibroblasts (i.e. 0% ASCs), its expression was only just detectable.
B: Comparison of cellular proliferation in SE with different amounts of ASCs (number of proliferating cells expressing Ki67 per field). The number of Ki67-positive cells, a proliferation marker, had significantly increased (p=0.05) when fibroblasts and ASCs were seeded at a 75% / 25% density with a value of 18 Ki67 positive-cells per field for this condition compared to a value of 6 Ki67 positive-cells per filed for fibroblast alone, whereas with a 50%/50% density of ASCs and fibroblasts alone, there was only a trend to increase at this culture time.
C: Comparison of epidermal thickness (µm) in SE with different amounts of ASCs. When DEs were seeded with cell suspension comprised 25% ASCs and 75% fibroblasts, the epidermal thickness was 3.2 times higher (with a value of 93,9 µm) than SEs prepared with 100% fibroblasts (with a value of 29,1 μm), whereas it was only 2.5 times higher with a DE prepared with 50% fibroblasts and 50% ASCs (with a value of 72,9 μm). Means calculated for 3 fields in 3 samples for each condition.
