Abstract
Rabbit conjunctiva, cornea, and iris have been used to evaluate and rank the ocular irritation potential of chemicals in the Draize eye irritancy test by subjective criteria. In order to rank the irritancy potential of chemicals objectively and biochemically, a primary culture method for rabbit conjunctival epithelial cells has been developed in our laboratory as a potential in vitro method for ocular toxicity testing of xenobiotics. Conjunctival epithelial cells were dispersed by Dispase II, followed by trypsin treatment. Cells were cultured in serum-free medium of 1:1 ratio of Dulbecco's modified Eagle's medium (DMEM) and F-12 nutrient mixture plus various concentrations of growth factors. At a plating density of 85,000 cells/cm2, cells grew to confluency in 3-4 days. Conjunctival cells showed a positive anti-keratin antibody stain which demonstrated their epithelial nature. These cells also showed positive periodic acid-Schiff (PAS) staining which is consistent with their goblet cell-containing and muc...
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