The invention belongs to the technical field of tissue engineering, and particularly relates to a method for inducing directed cartilage differentiation of stem cells. Based on a co-culture system ofcartilage cells and the stem cells, the cartilage cells and the stem cells are subjected to division and cooperation of labor through the co-culture system, an adenovirus vector is adopted to introduce a transforming growth factor (TGF-beta 3) into the stem cells, the continuous, local and over-expressed TGF-beta 3 are provided for the stem cells through transfected cartilage cells, directed cartilage cell differentiation of the stem cells is stimulated, exogenous addition of TGF-beta 3 is not needed, thus the growth factor is prevented from being decomposed by protease, damage possibly generated in the gene transfection process to the stem cells is avoided, and the higher survival rate and the better cartilage differentiation ability of the stem cells are maintained. In addition, throughthe mode of co-culture of the cartilage cells and the stem cells, directed cartilage differentiation of the stem cells in a cartilage cell conditional culture medium is facilitated, induction to the growth factor is acted more sensitively, and secretion of cartilage epimatrix is more facilitated.