The invention relates to the field of stem cell biology, specifically to a method for differentiating human pluripotent stem cells into natural killer cells and an application. The invention disclosesa pluripotent stem cell-derived natural killer cell. The pluripotent stem cell-derived natural killer cell expresses CD56, Nkp30, Nkp44 and Nkp46, and also expresses markers CD16 and CD94 of a maturenatural killer cell. The invention also discloses a method for preparing the natural killer cell. The method comprises the following steps: S1, formation of an embryoid body; S2, differentiation of the embryoid body into hematopoietic progenitor cells; S3, differentiation of the hematopoietic progenitor cells into NK cells; and S4, maturation and expansion of the NK cells. With the differentiation method provided by the invention, the pluripotent stem cells can be rapidly, efficiently, simply and conveniently induced to be differentiated into the natural killer cells with low cost on the basis of a culture medium with definite components and an optimized cell factor combination.