Culture method of breast cancer stem cells

The invention discloses a culture method of breast cancer stem cells. The method comprises the following steps of: adding obtained human breast cancer cells in a unicellular state into a complete culture medium to be made into a cell suspension; taking the logarithmic phase cell suspension, adding a proper amount of salmon thrombin, and after the uniform mixing, adding the same volume of salmon procollagen into the mixture to obtain a mixed solution; inoculating the mixed solution into a 96 pore plate of BME with the ultra-low adhesive force, and performing heat insulation at 36 to 38 DEG C so that the materials are solidified into soft gel; adding a DMEM/F12 culture solution, 1*B-27, epidermal growth factors, fibroblast growth factors, insulin-like growth factors-1 and hydrocortisone above the soft gel; and performing culture for 7 days to obtain breast cancer stem cell microspheres. The culture method has the advantages that the stem cell separation is stable; the effective amplification is realized; breast cancer stem cell microspheres are directly obtained; the microsphere cells have an ESA CD44 /CD24 phenotype; and the features of the tumor stem cells are obviously realized.