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.

[1]  A. Neufeld,et al.  Maintenance of corneal endothelial cell shape by prostaglandin E2: effects of EGF and indomethacin. , 1986, Investigative ophthalmology & visual science.

[2]  H. Kaufman,et al.  A medium-term corneal preserving medium (K-Sol). , 1986, Archives of ophthalmology.

[3]  A. Neufeld,et al.  Rabbit corneal endothelial cells in vitro: effects of EGF. , 1986, Investigative ophthalmology & visual science.

[4]  P. Binder,et al.  The growth of endothelium from human corneal rims in tissue culture. , 1984, Investigative ophthalmology & visual science.

[5]  E. Levine,et al.  Human endothelial cells: use of heparin in cloning and long-term serial cultivation. , 1983, Science.

[6]  R. Tripathi,et al.  Human trabecular endothelium, corneal endothelium, keratocytes, and scleral fibroblasts in primary cell culture. A comparative study of growth characteristics, morphology, and phagocytic activity by light and scanning electron microscopy. , 1982, Experimental eye research.

[7]  J. Baum,et al.  Studies of corneas in vivo and in vitro , 1981, Vision Research.

[8]  J. Wever,et al.  Effect of short- or long-term treatment with exogenous glycosaminoglycans on growth and glycosaminoglycan synthesis of human fibroblasts (WI-38) in culture , 1980, Mechanisms of Ageing and Development.

[9]  B. Yue,et al.  Mass culture of human corneal endothelial cells. , 1979, Archives of ophthalmology.

[10]  A. Mescher,et al.  Stimulation of corneal endothelial cell proliferations in vitro by fibroblast and epidermal growth factors. , 1977, Experimental eye research.

[11]  J. Baum,et al.  FINE STRUCTURE AND COLLAGEN SYNTHESIS ACTIVITY OF MONOLAYER CULTURES OF RABBIT CORNEAL ENDOTHELIUM , 1974, The Journal of cell biology.

[12]  J. Mannagh,et al.  Human corneal endothelium: growth in tissue cultures. , 1965, Archives of ophthalmology.

[13]  J. Quilligan,et al.  Growth Stimulating Effects of Acid Mucopolysaccharides.∗ , 1965, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.