The invention discloses an injection device for hair follicle stem cell culture and an application method of the injection device. The injection device for hair follicle stem cell culture used by themethod comprises storage structures, a first driving structure, a housing, a fixing structure, a storing structure, a second driving structure, a falling structure and a pressurizing structure. According to the injection device disclosed by the invention, the condition that the first driving structure is propped to be conveniently driven, is facilitated, so that the two storage structures are driven by the first driving structure to automatically open and close; the second driving structure for sealing the storing structure is arranged on the storing structure; the pressurizing structure for pressurizing digestive medicines is arranged in the storing structure; and the second driving structure is connected to the pressurizing structure, so that the condition that when the injection deviceis not used, the storing structure is sealed by the second driving structure, is facilitated; and when liquid medicines need to be used, the pressurizing structure is driven by the second driving structure for pressurizing to drive the liquid medicines in the storing structure to be drained, the using of the liquid medicines is more convenient and flexible, and besides, the pollution of the liquidmedicines is avoided.