Abstract The primary culture of rabbit corneal endothelium after isolation by trypsinization has been reported.1 Although this technique was uniformly successful in rabbits and monkeys, identical experiments upon human eyes failed with rare exception. A modification in technique has afforded success in 18 consecutive experiments on human corneas and, therefore, is thought worthy of reporting. This paper describes the modified technique for isolation and the primary growth of human corneal endothelium in tissue culture and presents observations of cell activity and the appearance of the endothelial mitochondria. A brief summary of the isolation and growth techniques which were unsuccessful is also reported. Materials and Methods The studies utilized human eyes which were not required for corneal transplantation. The eyes, varying from 28 to 70 years-of-age, were removed under surgically aseptic conditions within six hours after death, and washed with 0.5% ophthalmic Neosporin* solution. Cell culture was established 24 to 72 References 1. A solution of polymyxin B sulfate, neomycin sulfate, and gramicidin. 2. Slick, W.C.; Mannagh, J.; and Yuhasz, Z.: Enzymatic Removal and Pure Culture of Rabbit Corneal Endothelial Cells , Arch Ophthal 73:229, 1965.Crossref 3. Gwatkin, R.B.L., and Thompson, G.: New Method for Dispersing the Cells of Mammalian Tissues , Nature 201: 4925, 1964.Crossref 4. Polak, M.: Sobre una tecnica sencilla y rapida para la coloración del condrioma , Arch Histol 3:365-376, 1946.
Archives of Ophthalmology – American Medical Association
Published: Dec 1, 1965