The application discloses a method for separating and culturing primary
vascular smooth muscle cells from human abdominal
aortic wall, and relates to the technical field of
human cell culture. The method comprises the following steps: obtaining a
collagenase A solution; adding the
collagenase A solution to a treated human
abdominal aortic aneurysm wall
tissue sample and then
cutting the sample; transferring the
cut human
abdominal aortic aneurysm wall
tissue sample to a 37-degree
cell culture box for culture; centrifuging a
cell suspension obtained after termination of
digestion; discarding supernatant, resuspending
cell precipitate, and repeating
centrifugation for three times; resuspending the cell precipitate again, uniformly transferring the cell precipitate to a T25 culture
bottle after blowing, and placing the cell precipitate in a 37-degree
cell culture box for culture; when a wave peak and
wave trough state appears, the cell precipitate is digested and subcultured by using 0.25%
trypsin, and then the cell precipitate is transferred to a new T25 culture
bottle; and the cell precipitate is subcultured normally to the P3 generation and then stored in
liquid nitrogen, so that the
cell damage is prevented and the activity and proliferation capacity of the cell are maintained.