Electron microscopy of cultured human corneas. Osmotic hydration and the use of a dextran fraction (dextran T 500) in organ culture.

Human corneas were studied by means of electron microscopy after culture at 31 degrees C for two to 20 days in a medium containing 8% dextran T 500. Dextran T 500, a strong osmotic agent, was included in the culture medium to prevent excessive swelling of the cornea. In order to exclude the possibility that the observed effects were the result of osmotic changes during fixation, in each experiment, fixatives with different osmolalities were used (430, 574, 727, and 812 mOsm). After 8, 16, and 20 days of culture, vacuoles appeared that were filled with dextran; the cytoplasm was completely filled with these vacuoles. The vacuolization was not limited to the endothelium, but was also observed in stromal keratocytes and to a limited extent in the epithelium. It was concluded that the monolayer of endothelium was still intact after 20 days of culture in medium containing 8% dextran T 500, and, secondly, that the dextran might have been taken up by endocytosis. Whether or not the uptake of dextran has a long-term toxic effect on the corneal cells remains to be elucidated.