A three-dimensional polystyrene film was used to culture cell in vitro scaffolds, and the results were compared with those of the conventional two-dimensional liver cell culture.The proliferation and metabolism functions were analyzed, including the cell survival rate, albumin secretion, urea synthesis function and glucose consumption function.The results show that the three-dimensional cell proliferation activity is significantly higher than that of the two-dimensional and the gradually increased growth of the three-dimensional scaffold surface adhesion of liver cells and the formation of liver cell adhesion can be observed by biological microscope.The studies also show that the three-dimensional liver cells proliferation and metabolism functions are superior to the two-dimensional functions in the same conditions.The three-dimensional polystyrene film can be used as scaffolds of liver cell culture in vitro, and has good application prospect in the vitro cultivation system of biological artificial liver.