The invention discloses a method for separating sheep muscle stem cells. According to the method, muscle cells of the legs of a sheep are cut and obtained, the muscle cells are disinfected, cleaned, chopped, digested, centrifugally separated and the like, the muscle stem cells are separated and placed in a petri dish for culture; compared with conventional methods for separating the muscle stem cells, the method has the advantages that the separation efficiency is high, the infection rate in the separation process is low, the separated stem cells are high in survival rate and easy to culture,the number of the stem cells is increased, the muscle repair speed in the process of using the stem cells for treatment is increased, and the muscle repair efficiency in the process of using the stemcells for treatment is improved.