The invention discloses a culture medium for obtaining mesenchymal stem cells and exosomes thereof and a preparation method of the culture medium. DMEM / F12 is used as a basic culture medium, and the following components are added: 10-22 g / mL of a serum substitute, 20-55 ng / mL of a
growth promoting factor, 5-10 ng / mL of a
muscle cell repair factor HCGF, 5-16 mM of an anti-
aging factor, a buffer agent, 0.5-3 mg / L of an
antioxidant, 1-20 mg / L of vitamins,
antibiotics, 5-65 mg / L of trace elements and 1-16 [mu] g / mL of a protective agent. The
mesenchymal stem cell culture medium is definite in component, free of serum components, controllable in quality and capable of being produced on a large scale, stem cells can normally grow when the
mesenchymal stem cell culture medium is used for in-vitro culture of the stem cells, moreover, the
mesenchymal stem cell culture medium is beneficial to division and proliferation of the stem cells, the
stem cell amplification rate is greatly increased, and the mesenchymal
stem cell culture medium is suitable for large-scale production. Compared with the prior art, the
stem cell culture medium has the advantages that
stem cell culture time is shortened, and stem cells still can keep original biological characteristics and multidirectional differentiation potential after being subjected to multiple passage times by the aid of the
stem cell culture medium.