Abstract
Purpose: :
These studies were designed to evaluate the performance of the Clonetics® Human Corneal Epithelial Model in evaluating ocular barrier function and irritancy response.
Methods: :
The Clonetics Human Corneal Epithelial Model consists of primary human corneal epithelial cells, transfected with human papilloma virus (HPV) type 16 E6 and E7 genes, and grown on microporous membrane inserts. This culture system was evaluated for morphology and stratification using histological analysis. Barrier properties of the model system were characterized via sodium fluorescein permeability and by trans–epithelial electrical resistance (TEER). The cytotyoxicity response of the system to several known irritants was evaluated at concentrations correlating to a range of ocular irritancy categories using the MTT cytotoxicity assay.
Results: :
The observed histological profile of the culture model is reminiscent of that observed with in vivo corneal epithelium. Correspondingly, the system exhibited epithelial barrier properties consistent with the formation of intact epithelium. Lot–to–lot consistency in tissue response was demonstrated with statistical analysis of irritancy data for a 0.3% Triton X–100 control. The general irritancy response of the system to a panel of compounds showed good correlation with published Draize MMAS values.
Conclusions: :
Based on these results, the Clonetics Human Corneal Epithelial Model presents a good in vitro system for the assessment of ocular barrier function, toxicity and irritancy.
Keywords: cornea: epithelium • ocular irritancy/toxicity testing • cell adhesions/cell junctions