Antigen-specific T cell-derived induced pluripotent stem cells (iPSCs) have been shown to re-differentiate into functional T cells and thus provide a potential source of T cells that could be useful for cancer immunotherapy. Human Vα24⁺ invariant natural killer T (Vα24⁺iNKT) cells are subset of T cells that are characterized by the expression of an invariant Vα24-Jα18 paired with Vβ11, that recognize glycolipids, such as α-galactosylceramide (α-GalCer), presented by the MHC class I-like molecule CD1d. Vα24⁺iNKT cells capable of producing IFN-γ are reported to augment anti-tumor responses, which affects both NK cells and CD8⁺ cytotoxic T lymphocytes to eliminate MHC^- and MHC⁺ tumor cells, respectively. Here we describe a robust protocol to reprogram human Vα24⁺iNKT cells into iPSC, and then to re-differentiate them into Vα24⁺iNKT cells (iPS-Vα24⁺iNKT). We further provide a protocol to measure the activity of iPS-Vα24⁺iNKT cells.