The invention discloses an in-vitro culture solution of mesenchymal stem cells and a culture and amplification method. According to the in-vitro culture solution, DMEM culture solution is taken as a basic culture medium, compound (I), L-
glutamine,
vitamin C,
insulin,
erythrogenin and vascular
endothelial growth factors are sequentially added, then
sodium hydroxide is adopted for adjusting the pH, finally, fetal calf serum is added,
filtration sterilization is carried out, and thus the in-vitro culture solution of the mesenchymal stem cells is obtained, wherein the concentration of the compound (I) is 15-25 mg / L, the concentration of L-
glutamine is 200-400 mg / L, the concentration of
vitamin C is 60-90 mg / L, the concentration of
insulin is 10-20 mU / L, the concentration of the
erythrogenin is 20-40 U / L, and the concentration of the vascular
endothelial growth factors is 20-120 [mu] g / L. With the adoption of the in-vitro culture solution of mesenchymal stem cells and the culture and amplification method, the amplification efficiency of the human mesenchymal stem cells can be improved, and compared with the prior art, the in-vitro culture solution and the culture and amplification method have the outstanding substantive features and prominent progress.