The invention relates to a decellularization cornea preparation method, which adopts steps of corneal epithelium layer removing, ultraviolet cross-linking, viral inactivation, decellularization treatment, gradient dehydration, radiation protection and sterilization. According to the present invention, the prepared decellularization cornea has characteristics of complete antigen removing, no excitation of host acquired immunity reaction, good biocompatibility, low damage on nature corneal stroma, maintaining of structure characteristics of the nature corneal, and maintaining of effective components of the nature corneal so as to provide physical and chemical properties similar to the nature corneal; and after the prepared decellularization cornea is transplanted, animal experiment results show that characteristics of transparent transplanted cornea, no scar formation, no melting generation and no neovascularization are provided, the transplanted cornea and the recipient bed are completely integrated after transplanting a month, the transplanted cornea is subjected to complete epithelization after three months, corneal stroma cells migrate toward the decellularization cornea graft so as to prove that the tissue just takes place slow reconstruction, and the transplanted cornea after 6 months does not show significant difference in histology and appearance detection compared with the nature cornea.