Human corneal endothelium (HCE) is vital in maintaining corneal transparency and thickness, and damages to it may result in endotheliopathy and even corneal blindness, which can only be cured by keratoplasty. Due to the shortage of donor corneas, tissue-engineered HCE (TE-HCE) has become an equivalent to HCE. This study constructs a high endothelial cell density (ECD) TE-HCE and evaluates its functions by corneal transplantation in rabbits. A TE-HCE was constructed by culturing DiI-labeled nontransfected HCE cells on modified denuded amniotic membrane (mdAM) using collagen IV, fibronectin, and laminin, and rabbit corneas were characterized both in vivo and ex vivo after TE-HCE transplantation. Our results indicated that the constructed TE-HCE with a high ECD of 3611 ± 56.66 cells/mm2 had similar morphology and structure to a native HCE. In vivo postsurgery detections showed that the TE-HCE-transplanted cornea gained transparency and recovered its thickness gradually during a monitoring period of 358 days, while the mdAM-transplanted cornea remained opaque and edematous. Ex vivo examinations revealed that a native-like corneal endothelium with an ECD of 2703 ± 70.37 cells/mm2 was reconstructed by the transplanted TE-HCE. Our findings suggested that the TE-HCE might be used as a HCE equivalent and has promising applications in therapy of corneal endotheliopathy.