A method for inducing differentiated cells into mesenchymal stem cells (MSCs), and combinations of regulatory targets thereof. The method includes performing a directional induction on the differentiated cells to prepare the mesenchymal stem cells. The directional induction includes treating cells by inhibiting the TGF-β signal pathway, inhibiting the activity of PKC, activating the WNT/β-catenin signal pathway and activating the cAMP signal pathway. By regulating corresponding signal pathways and/or enzymatic activities by stages, the differentiated cells are induced into the mesenchymal stem cells.