The present invention relates to the technical field of
cell culture, and specifically discloses a method for domesticating an
adherent cell line into a fully suspended
cell line. The adherent-dependent cells are inoculated on microcarriers and
cell proliferation is carried out in DMEM medium containing 10%
fetal bovine serum. After the cells proliferate to 10E6 cells / mL, the microcarriers are lysed, the cell clusters are centrifuged and collected. The cell clusters are inoculated in a medium containing 10%
fetal bovine serum, 0.1%
shear force protectant, and 2 g /
L glucose and cultured at a rotation speed of 30 to 50 rpm for 3 to 4 days to enable the cells to form
embryoid body clusters of 50 μm to 150 μm. The
embryoid body clusters are mechanically pipetted to disperse them, and are domesticated for 18 to 22 days with DMEM medium containing different concentration gradients of
low molecular weight dextran sulfate to obtain a successfully suspended and domesticated cell line. The method of the present invention improves the uniformity of cell embryoid bodies, makes the properties of the embryoid bodies more stable, and can efficiently obtain a suspended cell line.