| abstract |
A process for establishing functional human corneal tissue consisting of an enhanced contact-inhibited endothelial monolayer derived from isolated human corneal endothelial cells with other integral corneal layers remaining intact. The process is divided into four integral parts. n 1. Isolation of human corneal endothelial cells from donor corneas. n 2. Establishment of a human corneal endothelial cell line which involves the establishment of primary cell cultures, proliferation, and continued maintenance and subculturing of these cells in vitro. n 3. The utilization of long term storage of isolated human corneal endothelial cells at -80° C. n 4. Utilizing these isolated corneal endothelial cells from the above processes to enhance the corneal endothelial monolayer of human donor corneas to be used for penetrating keratoplasty. n Two distinct methods of corneal endothelial enhancement may be utilized. In the first method, isolated human corneal endothelial cells are incorporated into the existing donor monolayer and re-establish a contact-inhibited functional monolayer, with a flattened, hexagonal configuration. In this procedure, the donor corneal endothelium remains intact and is supplemented with additional corneal endothelium. In the second method, isolated human corneal endothelial cells are seeded onto a denuded Descement's membrane and re-establish an intact cellular matrix, and form a contact-inhibited functional endothelial monolayer. In this procedure, the donor corneal endothelium is completely removed and replaced with new endothelial cells. |