The present invention relates to a method for isolating hair follicle stem cells and a composition for inducing hair growth. More specifically, relates to a method for isolating hair follicle stem cells showing a positive immunological response to CD34, by chemically degrading hair follicle-containing scalp tissue and then culturing the degraded tissue in a serum-containing medium and a serum-free medium, as well as a composition for inducing hair growth, which contains, as an active ingredient, CD34-positive hair follicle stem cells isolated by the method. The hair follicle-derived stem cells, which are obtained according to the disclosed method, are classified as autologous adult stem cells, have self-renewal capability, the ability to differentiate into adult hair follicle cells and the ability to induce hair growth, and can be used as a novel cell therapeutic agent against hair loss. In addition, the present invention relates to a method for culturing hair follicle cells, which has high yield compared to that of the prior art, as well as a method for identifying hair follicle stem cells.