The invention discloses a method for carrying out in-vitro efficient amplification on natural killer (NK) cells, and application of the method, aiming at solving the problems that the NK cells amplified by the traditional local method are small in amplification times, have cytokine dependency and cell aging, and the like. The method is characterized by comprising the following steps: 1, separating peripheral blood mononuclear cells (PBMCs) from human peripheral blood by using the traditional Ficoll-Paque density gradient centrifugation method; 2, carrying out 100 Grays irradiation treatment on artificial antigen presenting cells, and then storing by means of liquid nitrogen cryopreservation; 3, carrying out co-culture on the PBMCs and the artificial antigen presenting cells, subjected to the irradiation treatment, in a cell culture flask T75 of an RPMI 1640 medium, wherein the mass ratio of the PBMCs to the artificial antigen presenting cells is equal to 1: 2, and 50IU/mL of interleukin 2 is added into the RPMI 1640 medium; replacing the medium with a fresh medium every 2 to 3 days; 4, calculating the number of total cells in the cell culture flask T75 every 7 days, adding the same number of artificial antigen presenting cells subjected to the irradiation treatment into the cell culture flask T75 for stimulating again, and continuously culturing for 21 days to obtain the amplified NK cells. Therefore, after the method is adopted, a great deal of high-purity and high-quality NK cells can be obtained.