The invention relates to a method for constructing a human
corneal epithelial cell system. The construction method comprises the following steps of: disinfecting
cornea, digesting the
cornea, taking off
corneal epithelium with a front elastic layer,
cutting the
corneal epithelium into small tissue blocks, flatly attaching the tissue blocks to the bottom of culture holes downwards, adding culture solution into a
carbon dioxide culture box, and performing plate-attaching culture at the temperature of 37 DEG C, changing the culture solution during culturing, performing subculture when the cells grow into a single layer, and presently, completing the construction of the human
corneal epithelial cell system above 60 generations, wherein the culture solution contains 20 percent of fetal calf serum, 0.002 to 0.004 percent of human epidermic
cell growth factor, 0.001 to 0.002 percent of human alkali
fibroblast growth factor, 0.04 to 0.1 percent of carboxymethyl chito-
oligosaccharide, 0.025 to0.1 percent of DMEM / F12 culture solution of
chondroitin sulfate, and 0.008 to 0.01 percent of IV type collagen. In the construction method, the non-
transfection human
corneal epithelial cell system is successfully constructed by utilizing the human corneal epithelial tissues, and the constructed
cell system can realize continuous transfer of culture, and a great number of human corneal epithelialcells are provided.