The present invention relates to a method of inducing the differentiation of human pluripotent stem cells into endoderm and mesoderm by CXCR2 inhibition, and more particularly, to a method of promoting the differentiation of human pluripotent stem cells into endoderm and mesoderm by inhibiting the expression of the surface receptor CXCR2 in the stem cells by use of an shRNA technique. The method of inducing the differentiation of human pluripotent stem cells into endoderm or mesoderm by CXCR2 inhibition according to the present invention can increase the efficiency and utility of stem cells as a cell therapeutic agent, because it promotes the differentiation of stem cells into a specific germ layer serving as the origin of target cells, which is the first important step for inducing the differentiation of stem cells into specific cells.