The invention relates to a mesenchymal stem cell chemical component limited serum-free culture medium comprising a basic culture medium and additive components, and the additive components comprise: rhPDGF-BB, rhFGF-b, rhTGF-beta1, polyvinyl alcohol, alpha-ketoglutaric acid, lipopolysaccharide, recombinant fibronectin, recombinant human albumin, recombinant apo-transferrin, recombinant human insulin growth factors and hydrocortisone. The invention also provides application of the mesenchymal stem cell chemical component limited serum-free culture medium. The composition is clear and stable; instability of serum culture is eliminated, meanwhile, the factors have a synergistic effect, the adipose tissue-derived stromal cells can be efficiently amplified, the cell cycle is shortened, proliferation is rapid, the yield of the cells cultured for three generations reaches two times or above of that of serum culture, cell consistency is good, and the biological characteristics and differentiation potential of the cells can be efficiently maintained.