摘要Objective: Chimeric antigen receptor-modified T (CAR-T) cells have shown impressive results against relapsed/refractory B cell malignancies. However, the traditional manufacture of CAR-T cells requires leukapheresis to isolate large amounts of peripheral blood T cells, thus making some patients ineligible for the procedure.Results: The CAR-T cells were expanded to 1–3 × 108 cells in 8–10 d and successfully killed B cell-derived malignant tumor cells in vitro and in vivo. For patients with B-ALL, the complete remission rate was 93％ 1 month after CAR-T cell infusion; after 12 months, the overall survival (OS) and leukemia-free survival rates were 69％ and 31％, respectively. For patients with lymphoma, the objective response rate (including complete and partial remission) was 78％ 2 months after CAR-T cell infusion, and after 12 months, the OS and progression-free survival rates were 71％ and 43％, respectively. Cytokine-release syndrome (CRS) occurred in 65.51％ and 55.56％ of patients with B-ALL and B-cell lymphoma, respectively; severe CRS developed in 20.69％ of patients with B-ALL and in no patients with lymphoma.Conclusions: Our novel method can generate sufficient numbers of CAR-T cells for clinical use from 50–100 mL peripheral blood, thus providing an alternative means of CAR-T cell generation for patients ineligible for leukapheresis.