Figure 3.

Different growth by collagenase-isolated cells in coated, 2D, and 3D Matrigel. Single cells from collagenase-isolated limbal clusters (Fig. 1) were seeded in coated, 2D, and 3D Matrigel at 5 × 10⁴/cm² in MESCM. Spheres emerged in 3D Matrigel, whereas predominant spindle cells were found in coated and 2D Matrigel (A). The sphere in 3D Matrigel was formed by the reunion of single PCK+ (green) cells and Vim+ (red) cells, both of which increased in cell numbers in 10 days (B). Double staining between PCK (green) and EdU (red) confirmed that EdU+ nuclei were high in coated and 2D Matrigel but low in 3D Matrigel (C). Scale bar, 20 μm.