The invention relates to a method for preparing neural stem cells with high multiplication capacity, and belongs to the technical field of
biology. The method comprises the following steps: separating primary neural stem cells from
mammal embryo brain tissues, taking DMEM / F12 as a basic culture medium, adding B27, N2,
glutamine and double antibodies, and then adding epidermal growth factors, basic
fibroblast growth factors,
insulin-like
growth factor-1, Y-27632 and
heparin to form an optimized proliferation culture medium. The method comprises the following steps: inoculating cells into a culture dish coated with
polylysine and
laminin, culturing under proper conditions, and digesting and subculturing after neurospheres reach a certain
diameter; the method can improve the proliferation efficiency of neural stem cells and maintain stemness and differentiation potential, adopts a serum-
free system, is simple and convenient to operate, and is suitable for large-scale amplification and research application.