The invention relates to a method for efficiently separating and culturing mesenchymal stem cells of a primarily-cultured rabbit, which comprises the specific steps of: executing an
experimental animal by pumping
air embolism and then soaking for 15-20min with
alcohol for sterilizing, placing in a clean bench for separating
limb bones, shearing two ends of the bones after repeatedly washing with a PBS (
Phosphate Buffer) and DMEM (Dulbecco's Modified
Eagle Medium) culture solution, and washing marrow cavities with the DMEM solution containing
heparin with concentration of 625U / ml; respectively filtering marrow liquids obtained from the step 1 by using syringes with different diameters, preparing a monoplast suspension and then slowly injecting into an eccentric
pipe containing isovolumetric Percol 1 for centrifuging for 20min, collecting an intermediate cloud
cell layer, washing cells with an LG-DMEM solution, centrifuging for 5min, repeatedly washing for 2-3 times, adding the washed cells to the LG-DMEM solution containing a proper proportion of double antibodies (1%) and comprising 10% fetal calf serum, blowing and beating to form a
cell suspension, counting and regulating the
cell density to 1*10<8> / L, inoculating in a 25cm<2> plastic culture
bottle, and placing in a culturing box with conditions of a temperature of 37 DEG C and a
humidity of 5 percent for culturing. The invention can be effectively, conveniently and efficiently separating and culturing the mesenchymal stem cells of the primarily-cultured rabbit and is prepared for further basic experiments and
clinical research.