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Abstract
PURPOSE: To report on an in vitro reconstruction of cornea composed of three main layers. METHODS: These layers were reconstructed from isolated bovine corneal cells in three-dimensional collagen gel culture conditions. RESULTS: The topmost layer, corneal epithelial cells on collagen gel, was treated by what was designated as the "air-liquid interface method," a method developed specifically for this study. The treatment induced proliferation and differentiation of this layer. Those epithelial cells were positively immunostained with anti-epithelial keratin (55-kD/64-kD) antibody, AE5, which was specific for in vivo corneal epithelial cells. The middle layer consisted of keratocytes in the three-dimensional collagen gel matrix. This layer was comparable to the stroma in vivo. The underlayer, the counterpart of in vivo endothelial layer, was composed of corneal endothelial cells. CONCLUSIONS: This culture system probably provides a physiologic environment for detailed investigations of corneal cells.