The invention discloses a culture method of
breast cancer MCF-7
cell stem cell levitated spheres. The culture method comprises the following steps: 1) collecting
human breast cancer MCF-7 cells at a
logarithmic growth phase, paving the collected cells on a 75 to 83 percent culture
bottle, dissociating the cells with 0.25 percent by
mass pancreatin till the cells become round, and discarding the pancreatin; 2) adding a DMEM / H-G complete culture solution to end a reaction, blowing and beating with a dropper to form a single-
cell suspension, centrifuging at the speed of 1,000
revolutions per minute for 5 minutes, discarding supernatant, adjusting the
cell concentration, and performing passage in different bottles in the ratio of 1 to 3; 3) re-suspending the MCF-7 cells subjected to passage in a DMEM / F-12 culture medium which does not contain serum, adjusting the
cell concentration to 0.5*10<5> / mL, putting into a
penicillin glass
bottle, covering with a
bottle plug, forming an
oxygen-lack environment, culturing at the temperature of 37 DEG C in 5% CO2 without substituting the culture medium, mechanically blowing and beating into a single-cell suspension after a microcapsule is formed, and performing conventional passage. The
breast cancer MCF-7 cell
stem cell levitated spheres are cultured by an
oxygen-free culture method, and
spherical cell clusters remarkably growing in a suspending way appear on the second day of culture.