Process of endothelial cell-death in cultured corneas

Human and porcine corneas were cultured in organ-culture medium at 32 degrees C for long-term preservation and study of endothelial cell death. The endothelium was examined and re-examined at different intervals by using a phase contrast microscope and dual vital staining with alizarin-red S and trypan-blue. The appearance of degenerative vacuoles (which we called "death vacuoles" or "water-bubble bodies") as the first visible structural change in the process of cell death was very characteristic. While these vacuoles increased in size and number, the nucleus and cell-border disintegration was proceeding to the stage at which the cell was lost, leaving behind vacuoles on the Descemet's membrane, which we called the "endothelial graveyard."