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Figure 5: (a and b) Differentiation time course of dsASCs into epithelial-like cells on a collagen hydrogel showing (a) squamous cell-like morphology by day 4 and (b) epithelial-like cuboidal cell morphology by day 12. (c) Immunocytochemical image of section from dsASCs differentiated into epithelial-like cells on collagen gel (day 12) stained with pan cytokeratin. (d, e, g, and h) Differentiation time course of dsASCs in PEGylated-fibrin-collagen bilayer gels exhibiting bidirectional differentiation into fibroblast-like morphology in collagen layer (d; day 6) and tubular structures (g; day 6) in the PEGylated-fibrin layer. By day 12, collagen layers showed an increase in fibroblast-like cells (e) and complex networks in PEGylated-fibrin layers (h). Immunocytochemical image of section from bilayered hydrogel (day 12) depicting dsASCs to maintain stromal phenotype with collagen layer, stained with α-smooth muscle actin (f) and differentiated to vascular phenotype, stained with NG2 (i). (j and k) Differentiation time course of dsASCs into adipocytes on collagen showing appearance of oil vesicles by day 7 (j) and significant accumulation of oil droplets over time (k: day 14). (l) Oil Red O. staining of differentiated dsASCs (day 14) confirming formation of mature adipocytes within collagen gel. Bright field images original magnification: ×100 (a–h and l) and ×200 (j, k). Immunofluorescence original magnification ×400 (c, f, and i).