The invention discloses an Expi293F
cell culture method, which comprises the following steps: inoculating Expi293F cells into a
bioreactor for culture, and carrying out
perfusion dynamic culture for 32-48 hours: uniformly dividing a culture solution to be perfused into a plurality of strands, and perfusing into the
bioreactor along the inner wall of the
bioreactor in a wall-attached manner, with the stirring speed in the bioreactor being 80-90 rpm and the
perfusion rate of the perfused solution being 0.5-2 working volumes / d, during dynamic
perfusion culture, the culture conditions in the reactor are kept as follows: the temperature is 36.5-37 DEG C, the DO is 40-60%, and the pH is 7.2 + / -0.2; a serum-free culture solution used in the culture process is prepared from 95wt% of 24.75 g / L Dynami culture medium, 4wt% of 200mM
glutamine and 1wt% of an anti-
caking agent. According to the invention, the perfusion mode is improved, and proper perfusion rate and stirring
strength reduction are matched, so that the injection of the culture medium in the perfusion process also plays roles in expanding
mass transfer, reducing the influence of
cell agglomeration on
cell metabolism and reducing physical damage to cells, high-density cells and
high survival rate are obtained, and remarkable effects are achieved.