The efficient infiltration of immune cells into tumors is a prerequisite for effective cancer immunotherapy. Eosinophils have been demonstrated to be able to quickly infiltrate tumors and recruit other immune effector cells to enhance the effects of antitumor treatments, indicating that eosinophils might provide the key to more effective cancer immunotherapies. In this study, we established a stepwise differentiation approach for generating large numbers of functional EPX+ eosinophils from human pluripotent stem cells (hPSCs) at near-100% purity. These hPSC-derived cells not only displayed eosinophil-specific phenotypes and expressed cytotoxic granular proteins, but more importantly, could also kill tumor cells in vitro, quickly infiltrate solid tumors in vivo and significantly suppress tumor growth in colorectal carcinoma and ovarian cancer xenograft mouse models. Our study therefore opens new avenues for the development of eosinophil-based immunotherapies to treat cancer. Overall design: our goal was to compare and analysis the transcriptome of eosinophils from PSC with human naïve eosinophils from Cord blood. This series includes tumor samples from cancer xenograft mice injected with CAR-T cells and tumor samples from cancer xenograft mice injected with the combination of CAR-T cells and PSC-derived eosinophils (E).