PURPOSE. To develop and evaluate a three-dimensional organ culture system of the cornea anterior chamber that could replicate the in vivo processes occurring during corneal wound healing and corneal transplantation. METHODS. Bovine corneoscleral buttons were clamped in a specially designed chamber through the sclera outside the limbus. The epithelium was exposed to air, and its anterior surface was automatically irrigated. The endothelial layer was perfused separately with media under normal intraocular pressure. Wound healing and corneal transplantation were observed using light, scanning, and transmission electron microscopy. RESULTS. The organ culture system maintained the epithelium, the putative epithelial stem cells in the limbus, the stroma, and the endothelium in good condition for the 10-day period during which the system was evaluated. The authors observed that the processes of wound healing and corneal transplantation in the model appeared similar to those occurring in vivo. CONCLUSIONS. In vitro model closely replicated the in vivo processes of wound healing and corneal transplantation. The authors believe this model will be useful for basic investigations into the cornea, such as study of the response of the cornea to surgery, wound healing, toxins, and therapeutic agents.