A Human Corneal Epithelial Culture Model

Abstract

Abstract: : Purpose: We have developed an in vitro culture model comprising human primary corneal epithelial cells in a physiologically relevant environment. The system can be used as a research tool to assess ocular toxicity and irritancy Methods: Human corneal epithelial cells were isolated and transfected with human papilloma virus (HPV) type 16 E6 and E7 genes. Culture models were developed by growing the cells to confluence on microporous membrane supports in Corneal Epithelial Model Medium followed by culture at the air–liquid interface to induce stratification and differentiation of the cells. Cultures were assessed for differentiated function via Trans Epithelial Electrical Resistance (TEER), histological analysis, and time–to–toxicity (ET₅₀) as assessed by the MTT cytotoxicity assay for several known irritants. Results: The culture models exhibited a histological profile similar to that observed with in vivo corneal epithelium. Furthermore, the cytotoxic response of the system to known irritants shows good correlation with in vivo Draize data for several different irritant categories (severe, moderate, mild, slight and non–irritants). Conclusions: Based on these results, the human corneal epithelial culture model described here presents a promising in vitro system for the assessment of ocular toxicity and irritancy.

Keywords: ocular irritancy/toxicity testing • cornea: epithelium • drug toxicity/drug effects