[Growing human corneal endothelium in cell culture].

Enzyme treatment of the corneal endothelium with collagenase was employed in a new method of isolating human corneal endothelial cells (HCEC) from the corneas of adult donors (30-70 years). It was possible to isolate 5-10 X 10(4) endothelial cells from each cornea, and primary cultures were established. During the first 3-5 passages, the growth of contaminating fibroblast like cells was inhibited by the use of an L-valine-free selective culture medium. Proliferation of the cells was stimulated by supplementation of the culture medium with fibroblast growth factor and also by coating the culture dishes with a mixture of laminin and chondroitin sulfate. After 3-6 passages, the cell number had increased 150-200 times. During the passage the cells changed with regard to their typical morphology. This change depended on the amounts of serum and mitogen, respectively, in the culture medium, as well as on the in vitro age of the cells.