The invention provides a method for separating and culturing olfactory ensheathing cells of mice. The method comprises the steps of taking a female mouse in the middle and late pregnancy, taking out amouse embryo, taking out an olfactory bulb of the mouse embryo, cutting the olfactory bulb tissue into pieces, adding a complete medium containing serum, filtering with a 200-mesh sieve, sucking thetissue fluid out and adding into a centrifuge tube for centrifuging, discarding the supernatant, resuspending, and repeating the operation for three times; adding into the complete medium to adjust the cell concentration, and culturing in a constant temperature incubator; then, using a differential adhension method to purify the olfactory ensheathing cells. After the method for separating and culturing the olfactory ensheathing cells of the mice is adopted, the separated and cultured olfactory ensheathing cells are large in quality, high in density and strong in biological activity.