The manufacturing of autologous chimaeric antigen receptor (CAR) T cells largely relies either on fed-batch and manual processes that often lack environmental monitoring and control or on bioreactors that cannot be easily scaled out to meet patient demands. Here we show that human primary T cells can be activated, transduced and expanded to high densities in a 2 ml automated closed-system microfluidic bioreactor to produce viable anti-CD19 CAR T cells (specifically, more than 60 million CAR T cells from donor cells derived from patients with lymphoma and more than 200 million CAR T cells from healthy donors). The in vitro secretion of cytokines, the short-term cytotoxic activity and the long-term persistence and proliferation of the cell products, as well as their in vivo anti-leukaemic activity, were comparable to those of T cells produced in a gas-permeable well. The manufacturing-process intensification enabled by the miniaturized perfusable bioreactor may facilitate the analysis of the growth and metabolic states of CAR T cells during ex vivo culture, the high-throughput optimization of cell-manufacturing processes and the scale out of cell-therapy manufacturing.

自体嵌合抗原受体 （CAR） T 细胞的生产在很大程度上依赖于通常缺乏环境监测和控制的补料分批和手动工艺，或者依赖于无法轻松扩展以满足患者需求的生物反应器。在这里，我们表明，人原代 T 细胞可以在 2 ml 自动闭式系统微流控生物反应器中被激活、转导和扩增至高密度，以产生可行的抗 CD19 CAR T 细胞（具体来说，超过 6000 万个来自淋巴瘤患者供体细胞的 CAR T 细胞和超过 2 亿个来自健康供体的 CAR T 细胞）。细胞因子的体外分泌、细胞产物的短期细胞毒活性和长期持久性和增殖，以及它们的体内抗白血病活性，与在透气井中产生的 T 细胞相当。小型化可灌注生物反应器实现的制造过程强化有助于分析离体培养过程中 CAR T 细胞的生长和代谢状态、细胞制造过程的高通量优化和细胞治疗生产的规模化。