In recent years cancer immunotherapy， especially the cell-based immunotherapy， has reached several milestones and achieved a lot of cancer remission in the clinics. Obtaining a more potent and effective cytotoxic T lymphocytes (CTLs) for cancer immunotherapy is always the ultimate goal for the researchers. However， the difficulty in harvesting a large number of tumor antigen-specific CTLs from the tumor patient is still a major obstacle we need to overcome. In our previous studies， it is shown that pluripotent stem cell-derived CTL-especially the genetically engineered antigen-specific CTLs-may serve as a good source of unlimited number of highly reactive and antigen-specific CTLs. Here we present a two-step method for the generation of antigen-specific T lymphocytes from iPS cells by in vitro priming and in vivo maturation.