The invention provides an induction composition of human mesenchymal stem cells, an induction differentiation culture liquid and an in-vitro induction method and an application, and relates to the field of nerve cell regeneration. The induction composition includes a brain-derived neurotrophic factor, a nerve growth factor, an epidermal cell growth factor, a fibroblast growth factor, a N2 cell culture additive, a B27 cell culture additive, rhodioloside and astragalus polysaccharide. In the induction composition provided by the invention, rhodioloside, astragalus polysaccharide and cytokines are compounded, induced differentiation nerve cells have the advantages of high content, strong activity, short period and low cost, can solve the problem of low differentiation efficiency when human mesenchymal stem cells are induced in vitro, improve the survival status of the induced nerve cells, and have little toxic and side effects on cells. The induction composition can be used for inducing the human mesenchymal stem cells, is easy to operate, enhances the activity of the differentiated nerve cells and shortens the induction differentiation time.