The use of an in vitro cornea for predicting ocular toxicity

This paper presents the development of an in vitro cornea model to assess ocular irritancy. The in vitro cornea is an organotypic culture composed of corneal stromal fibroblasts embedded in a matrix of collagen with an underlying layer of endothelial cells and covered by corneal epithelial cells. All corneal cells were isolated from fetal pig. Pure cultures of the three different cell types were obtained using an improved technique for isolation and cultivation. Subcultures were passaged until the 30th generation without showing signs of senescence. For a cytotoxicity assay in vitro corneas were constituted in microtiter plates. The cytotoxic response of five test chemicals with a well-characterized eye irritancy was studied using a modified MTT-assay, which possess many advantages over the traditional procedure. The irritation data for all test chemicals yieled by our test assay correlated well with the Draize test classification.