Fig. 4
From: Sphere-forming cells from peripheral cornea demonstrate the ability to repopulate the ocular surface
Implantation of peripheral corneal spheres into corneoscleral tissue results in cell migration, inter-sphere interaction and polarized outgrowth. LIVE/DEAD® staining of sphere-implanted tissue showed a green (live cell) fluorescent signal in the outline of the sphere with the beginnings of cell migration and minimal tissue staining (arrows) at 25 h a. An increase in live cell migration is observed in tissue over time from 72 h b to 217 h c. Arrow indicates a blurred region of tissue staining in a different plane of focus c. White lines at all three time points are of equal lengths showing a decline in sphere diameter from 72 h b to 217 h c. In cross-section, cell nuclei labelled using DAPI (blue) are dispersed d. Confocal imaging of EDU staining shows red signals in the sphere (arrow) and in migrated cell nuclei (arrowhead) e. Polarized cell migration is evidenced in implanted spheres f. Two spheres implanted adjacent to each other in tissue are imaged first at 72 h g, g1 and subsequently at 217 h h. Here, cells migrate from each sphere in the direction of each other. Migration on the right of the diagonal line appears to have increased from 72 h to 217 h more so than on the left of the diagonal line. Montage imaging of the spheres at 72 h post implantation with light microscope image overlay shows the position of spheres at the limbal region of corneoscleral rims g1. Scale bar = 100 μm (Colour figure online)