Reconstruction of cornea in three-dimensional collagen gel matrix culture.

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.

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