Optimal approaches to differentiate tumor antigen-specific cytotoxic T lymphocytes (CTLs) from pluripotent stem cells (PSCs) remain elusive. In the current study， we showed that combination of priming through Notch ligands and development facilitated the generation of tumor Ag-specific CTLs that effectively inhibited tumor growth. We co-cultured the murine induced PSCs (iPSCs) genetically modified with tyrosinase-related protein 2 (TRP2)-specific T cell receptors with OP9 cell line expressing both Notch ligands Delta-like 1 and 4 (OP9-DL1/DL4) for a week before adoptively transferred into recipient C67BL/6 mice. Three weeks later， B16 melanoma cells were inoculated subcutaneously， and the antitumor activity of the iPSC-derived T cells was assessed. We observed the development of the TRP2-specific iPSC-CD8 T cells that responded to Ag stimulation and infiltrated into melanoma tissues， significantly inhibited the tumor growth， and improved the survival of the tumor-bearing mice. Thus， this approach may provide a novel effective strategy to treatment of malignant tumors.