Preclinical testing of small diameter Descemet membrane endothelial keratoplasty grafts to increase tissue availability
Fig 5
Example of in vitro endothelial cell migration from a gel-cultured 4mm graft.
(A) Light microscopy overview collage (50x magnification) of a 4 mm DMEK graft after 17 days in gel culture showing uniform cell migration from all around the graft. (B) Light microscopy image showing central graft endothelium and (C) graft edge (bottom of the image) from where migration was directed as a confluent cellular monolayer. (D, E) Fluorescence microscopy images showing expression of ZO-1 (red signal) counterstained with DAPI (blue signal) in the graft center and in the migrated monolayer. The absence of ZO-1 stained cell borders in the lower part of image 5E can be attributed to the fact that cells in this area reside on the graft and are elevated as compared to the new cell monolayer. Therefore, the cells on the graft have an elevation of about 10 μm (on the Descemet membrane) when compared to the migrated cells on the glass cover slide and therefore appear out of focus. Scale bars: 100 μm.