An articular cartilage graft and a preparation method thereof are provided. The prepared articular cartilage graft is composed of a superficial layer, a middle layer and a deep layer from outside to inside; the thickness, shape and size of each layer are all matched with a cartilage injury part, and thus preoperative shaping is not required; after grafting, the articular cartilage graft has layer distribution corresponding to distribution of each layer of surrounding normal cartilages, is conducive to intercellular signal transmission, transduction and regulation, and can be better integrated with surrounding normal cartilage tissues; the articular cartilage graft has structure characteristics consistent with those of the natural cartilages, the collagen type II content is gradually decreased from the superficial layer to the deep layer, the GAG content is increased gradually from the superficial layer to the deep layer, and compression resistance and wear resistance are good; and chondrocytes in the cartilage graft are wrapped with an extracellular matrix, have low immunogenicity, allow generation of immunologic rejection to be avoided after grafting, can survive for a long term and exert functions, and improve cartilage repair long-term curative effects.