The invention discloses a primary isolation culture method of adipose-
derived stem cells. The primary isolation culture method comprises the following steps: (1) washing adipose tissues; (2) adding a mixed liquid of type-I
collagenase and trypsase into the adipose tissues, wherein the volume of the mixed liquid is equal to the volumes of the adipose tissues; (3) adding FBS, covering a
bottle cap, sealing, uniformly shaking, carrying out
centrifugation, and discarding supernate after the
centrifugation; (4) carrying out resuspension
precipitation by virtue of a DMEM containing the FBS, uniformly blow-hitting to form
cell suspension, and inoculating the
cell suspension to a culture medium, wherein the volume ratio of precipitates to the DMEM containing the FBS is 1 to 1; (5) culturing the inoculated culture medium; and (6) transferring the cultured culturing medium into a
cell incubator, and continuing to culture the culturing medium. Enzymolysis conditions explored in the primary isolation culture method are stable and efficient, the complete enzymolysis of the adipose tissues is achieved, and cells of each component are furthest released; on the premise that the activity of ADSCs is not influenced, the quantity of the ADSCs acquired in unit adipose volume is relatively stable.