97th DOG Annual Meeting 1999
CHARACTERISATION OF THREE DIFFERENT CELL TYPES IN HUMAN AND PORCINE CORNEA
K. Gaebert1, E. Berger1, J. Saedler1, D. G. Weiss2,
Purpose: The cornea is subdivided in three different layers: epithelium, stroma and endothelium. The first step for the development of an artificial cornea is the characterisation of these cell types. Corneal stromal fibroblasts are the most important cells to anchor the keratoprosthesis to the host tissue.
Materials and Methods: Corneal epithelial, stroma and endothelial cells were isolated from porcine eyes according to the method described by Schneider et al. (In Vitro Toxicology 10(3), 1997, 309-18). For preparation of human corneal fibroblasts we developed a method with a special cell culture medium. The differentiation of the cell types was made by morphological determinations and immunocytochemical detection of cytoskeletal filaments and membrane proteins.
Results: Vimentin could be detected in endothelial and stroma, but not in epithelial cells. To differentiate both cell types an Anti-Human Fibroblast Surface protein was used and could be detected only in stroma cells. There was no detection of desmin and cytokeratin in one of these three cell types.
Conclusion: We were able to cultivate and characterise the corneal epithelial, stromal and endothelial cells. The identification of keratocytes of human adult eyes was possible through the localisation of the intermediary filament vimentin and the fibroblast surface protein by immunofluorescence microscopy.
1Dept. of Ophthalmology, University of Rostock, Doberaner-Str. 140,
D- 18057 Rostock, Germany
2Dept. of Biology, University of Rostock, Universitätsplatz 2,
D-18057 Rostock, Germany