The present invention relates to a method for inducing amplification of human type I NKT cells in vitro using a “specific stimulant+staged cytokine” mode, which consists of two culture stages, wherein the first culture stage focuses on specific amplification of the number of the type I NKT cells, in which a specific stimulant α-GalCer is used to advantageously amplify the type I NKT cells and α-GalCer-loaded CD1d-expressing cells are used to stimulate continuous proliferation of the type I NKT cells while adding cytokines IL-2 and IL-7 to assist growth of the type I NKT cells; and the second culture stage is to synchronously perform amplification of the number of the type I NKT cells and guide directed function differentiation, in which CD1d-expressing cells incubated with α-GalCer continue to stimulate proliferation of the type I NKT cells while adding IL-2, IL-7 and IL-15 to assist amplification of the type I NKT cells and guide differentiation, and IL-12 is added to the culture system 1-2 days before the end of culture to guide further directed differentiation of the type I NKT cells and enhance their killing activity. The method of the present invention is simple to operate, can greatly amplify the type I NKT cells in vitro while improving the killing activity of the amplification products, and is suitable for large-scale production.