Corneal endothelial replacement. I. In vitro formation of an endothelial monolayer.

Cultured bovine corneal endothelial cells were seeded onto a corneal button denuded of its own endothelium. After an incubation time of 30 min to 1 hr, these cultured cells were able to repopulate and reconstitute a new endothelium over Descemet's membrane. Three basic cellular processes were involved in the formation of the new endothelial layer. (1) Adhesion of the cells to the tissue substrate occurred rapidly because Descemet's membrane is the natural substrate for these cells. (2) Expansion of the cells formed a continuous monolayer and, at the concentration used in our experiments, was achieved with a minimum of cell thinning. (3) Cohesion was developed within 1 hr after seeding when cells contacted each other by means of long thin cytoplasmic processes (filopodia). The special junctional complexes associated wih the fluid barrier found in the corneal endothelium were only partially formed 72 hr after plating the cultured cells. Finally, the durability of the reconstituted endothelium was examined by using the corneal buttons at keratoplasty. It appears that corneal buttons incubated for 1 to 2 hr may be used at keratoplasty.