Growth of human corneal endothelial cells in culture

We investigated the effects of various culture conditions on the growth of normal human corneal endothelial cells in culture. Falcon Primaria tissue culture plastic was found to provide a more suitable surface for endothelial cell growth than the conventional Corning tissue culture plastic. Also, media containing 10% fetal bovine serum and 5% calf serum (complete media) facilitated the growth of human cells better than those containing Nu-serum. Supplementation with epidermal or fibroblast growth factor (10 and 100 ng/ml) to the complete media had no effect on human endothelial cell growth. Chondroitin sulfate at low concentrations (100 micrograms/ml to 1 mg/ml) also showed little effect. At high concentrations (13.5 and 25 mg/ml), however, chondroitin sulfate significantly promoted human corneal endothelial cell growth during a 1- to 2-week incubation period. From the 37 cultures initiated, outgrowth from explants appeared within 3 to 7 days. Cells were polygonal in shape and, at confluency, formed a continuous monolayer. We attained a success rate of 87% (7/8) growing primary cultures from donors under 20 years of age and a 59% (17/29) success rate from older donors.