A clinically guided method has been developed to generate allogeneic CAR-NKT cells from hematopoietic stem and progenitor cells (HSPCs), enabling high-yield and high-purity production without the need for three-dimensional culture or xenogeneic feeders. These cells, engineered to target multiple cancers, demonstrated potent antitumor efficacy in a multiple myeloma model, with enhanced persistence and selective depletion of immunosuppressive cells in the tumor microenvironment. AlloCAR-NKT cells exhibit a stable hypoimmunogenic phenotype, regulated by epigenetic and signaling mechanisms, and do not induce graft-versus-host disease or cytokine release syndrome. They effectively target tumors through CAR, TCR, and NK receptor mechanisms, showing superior antitumor activity compared to conventional CAR-T cells. AlloCAR-NKT cells also display enhanced NK features, making them particularly effective against solid tumors. The cells were generated from cord-blood HSPCs using a five-stage, 6-week process, with high expansion rates and purity. Preclinical studies confirmed their safety, efficacy, and ability to persist long-term. AlloCAR-NKT cells show promise for clinical translation due to their off-the-shelf potential, reduced risk of GvHD, and broad therapeutic applications. The study highlights the potential of allogeneic CAR-NKT cells as a promising alternative to autologous CAR-T cells in cancer immunotherapy.A clinically guided method has been developed to generate allogeneic CAR-NKT cells from hematopoietic stem and progenitor cells (HSPCs), enabling high-yield and high-purity production without the need for three-dimensional culture or xenogeneic feeders. These cells, engineered to target multiple cancers, demonstrated potent antitumor efficacy in a multiple myeloma model, with enhanced persistence and selective depletion of immunosuppressive cells in the tumor microenvironment. AlloCAR-NKT cells exhibit a stable hypoimmunogenic phenotype, regulated by epigenetic and signaling mechanisms, and do not induce graft-versus-host disease or cytokine release syndrome. They effectively target tumors through CAR, TCR, and NK receptor mechanisms, showing superior antitumor activity compared to conventional CAR-T cells. AlloCAR-NKT cells also display enhanced NK features, making them particularly effective against solid tumors. The cells were generated from cord-blood HSPCs using a five-stage, 6-week process, with high expansion rates and purity. Preclinical studies confirmed their safety, efficacy, and ability to persist long-term. AlloCAR-NKT cells show promise for clinical translation due to their off-the-shelf potential, reduced risk of GvHD, and broad therapeutic applications. The study highlights the potential of allogeneic CAR-NKT cells as a promising alternative to autologous CAR-T cells in cancer immunotherapy.