Protocol to generate human stem cell-derived CD70-directed allogeneic CAR-NKT cells for treating renal cell carcinoma.

CD70 has emerged as a promising immunotherapeutic target in renal cell carcinoma (RCC), with both CD70-directed monoclonal antibodies and chimeric antigen receptor (CAR)-based therapies currently under development. Here, we describe a protocol for the generation of human CD70-directed allogeneic CAR-natural killer T (NKT) ((Allo)CAR70-NKT) cells derived from cord blood CD34(+) hematopoietic stem and progenitor cells (HSPCs) using a clinically guided culture. Furthermore, we describe the therapeutic efficacy of (Allo)CAR70-NKT cells in mediating cytotoxic activity against RCC cell lines in vitro. For complete details on the use and execution of this protocol, please refer to Li et al.(1).