Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, London, WC1E 6BT, UK.
Aston Medical Research Institute, School of Life and Health Sciences, Aston University, Birmingham, B4 7ET, UK.
Biotechnol J. 2020 Sep;15(9):e2000177. doi: 10.1002/biot.202000177. Epub 2020 Jul 12.
Chimeric antigen receptor T-cell (CAR-T) therapies have proven clinical efficacy for the treatment of hematological malignancies. However, CAR-T cell therapies are prohibitively expensive to manufacture. The authors demonstrate the manufacture of human CAR-T cells from multiple donors in an automated stirred-tank bioreactor. The authors successfully produced functional human CAR-T cells from multiple donors under dynamic conditions in a stirred-tank bioreactor, resulting in overall cell yields which were significantly better than in static T-flask culture. At agitation speeds of 200 rpm and greater (up to 500 rpm), the CAR-T cells are able to proliferate effectively, reaching viable cell densities of >5 × 10 cells ml-1 over 7 days. This is comparable with current expansion systems and significantly better than static expansion platforms (T-flasks and gas-permeable culture bags). Importantly, engineered T-cells post-expansion retained expression of the CAR gene and retained their cytolytic function even when grown at the highest agitation intensity. This proves that power inputs used in this study do not affect cell efficacy to target and kill the leukemia cells. This is the first demonstration of human CAR-T cell manufacture in stirred-tank bioreactors and the findings present significant implications and opportunities for larger-scale allogeneic CAR-T production.
嵌合抗原受体 T 细胞（CAR-T）疗法已被证明在治疗血液系统恶性肿瘤方面具有临床疗效。然而，CAR-T 细胞疗法的生产成本非常高。作者在自动化搅拌罐生物反应器中展示了从多个供体制造人 CAR-T 细胞的方法。作者在搅拌罐生物反应器中成功地在动态条件下从多个供体中生产出功能性人 CAR-T 细胞，从而获得的总体细胞产量明显优于静态 T 瓶培养。在搅拌速度为 200rpm 及以上（高达 500rpm）的情况下，CAR-T 细胞能够有效增殖，在 7 天内达到超过 5×10^6 个细胞/ml 的活细胞密度。这与目前的扩增系统相当，明显优于静态扩增平台（T 瓶和气相通培养袋）。重要的是，经过扩增后的工程化 T 细胞保留了 CAR 基因的表达，并保留了其细胞溶解功能，即使在最高搅拌强度下生长也是如此。这证明了本研究中使用的功率输入不会影响靶向和杀死白血病细胞的细胞功效。这是首次在搅拌罐生物反应器中制造人 CAR-T 细胞的演示，这一发现为更大规模的同种异体 CAR-T 生产带来了重要的意义和机遇。
