Figure 2.

Cell sheet formation and identification. (A) Cell sheet formation after 21 days of continuous culture. The first line shows photographs of each of the types of cell sheets; the second line show HE staining results of three cell sheets, which revealed that the cultured oral mucosal epithelial cell sheets were composed of 2-3 layers of cells (left, scale bar: 50μm), the cultured oral fibroblast cell sheets were composed of 3-4 layers of cells (middle, scale bar: 100μm), and the cultured myoblast induction of ADSCs sheets were composed of 6-7 layers of cells(right, scale bar: 100μm); the third line shows the SEM images of the three cell sheets, scale bar: 50 μm. (B) Immunohistochemical identification of three types of cell sheets. The result of immunohistochemistry revealed, positive staining of pan cytokeratin (AE1+AE3) in oral mucosal epithelial cell sheets (a), b is the negative control; positive expressions of muscle protein markers in myoblast induction of ADSCs sheets after 3 weeks of culture with myoblast differentiation medium, including α-SMA (c), PAX7 (d) and Desmin (e), f is the negative control; positive staining of vimentin in oral fibroblast cell sheets (g), h is the negative control. Scale bar: 100 μm.