Abstract: Fibroblast (FB) has shown unprecedented potential in cosmetic and regenerative medicine. However, the limited availability and quantity of fibroblasts are bottlenecks limiting their clinical application. In this study, we use DFM, a fibroblast-induced differentiation medium designed by our laboratory, to induce the differentiation of Mesenchymal Stem Cells (MSCs) into fibroblasts in vitro. The cell proliferation characteristics after induction are determined by cell counting, the expression of fibroblast-related genes is detected using qRT-PCR, and the expression of fibroblast-related proteins is measured by flow cytometry. In addition, the migration ability of induced cells is determined by scratch tests. Based on these data, a comprehensive evaluation system for cell differentiation efficiency is established. The results show that the expansion folds of total cells in the induction medium DFM are significantly higher than those in the control group; The expression of fibroblast-related genes such as VIM (vimentin), LN (laminin), and FSP-1 (fibroblast-specific protein 1), as well as the expression of fibroblast-related proteins (including VIM, LN, and FSP-1) are significantly up-regulated in comparison with those in the control group. The migration ability of induced cells is also significantly enhanced in comparison with the control group.