The present invention is a method, belonging to the field of cell culture, for amplifying a large numbers of hair follicle stem cells in vitro by using microspheres as carriers for cell culture and a revolving bottle as a fermentation tank for cell proliferation. The method is simple in its procedures, is economical and practical, and avoids side effects associated with the traditional cell amplification method, such as the weakening of cell proliferation capability, the reduction of differentiation potential and the like caused by repeated cell subculture, and also reduces the consumption of the culture solution; the hair follicle stem cells amplified by the method can still keep the original proliferation capability and differentiation potential, and can be used for: (1) establishing hair follicle stem cell bank to provide high-quality seed cells for related research on adult stem cells; (2) transplanting and repairing tissues and organs subjected to pathological changes; (3) constructing tissue-engineered organs to be used as substitute autologous organs for transplantation or repair of organs subjected to pathological changes or missing organs; and (4) serving as target cells for the genetic treatment of corresponding diseases.
