The invention provides a method for orient induction amplification of thymus gland-derived iNKT (Invariant Natural Killer) cells. The method comprises two stages, namely a first stage, including stepsof performing in-vivo stimulation of iNKT cell proliferation, namely stimulating in-vivo iNKT cell proliferation of mice by using a specific stimulant alpha-Galcer; a second stage, including steps ofperforming in-vitro orient induction of iNKT cell proliferation, namely stimulating thymus glands of the mice by using alpha-Galcer, preparing a single-cell suspension, adding alpha-Galcer and an induction agent, assisting iNKT cell amplification, and inducing cell differentiation. With the combination of in-vivo proliferation stimulation and in-vitro induction amplification, orient iNKT cell induction is achieved, and the purpose that a great amount of orient differentiation iNKT cells with specific functions are obtained within a short time is achieved. Complex and tedious steps of in-vitrocell amplification culture are effectively reduced, the cell culture time is shortened, good repeatability is achieved, and the cost is relatively low.