Matsumoto Masahiro, Tashiro Shinji, Ito Tatsumi, Takahashi Kazuya, Hashimoto Gakuji, Kajihara Junji, Miyahara Yoshihiro, Shiku Hiroshi, Katsumoto Yoichi
Tokyo Laboratory 11, R&D Center, Sony Group Corporation, Tokyo, Japan.
Department of Personalized Cancer Immunotherapy, Mie University Graduate School of Medicine, Mie, Japan.
Mol Ther Methods Clin Dev. 2023 Aug 2;30:367-376. doi: 10.1016/j.omtm.2023.07.012. eCollection 2023 Sep 14.
By analyzing patients treated with adoptive immune cell therapies, various immune cell phenotypes have been found in the starting and infused materials as determinants of sustained remission. The isolation of these specific phenotypes for clinical use requires current Good Manufacturing Practice (cGMP)-compliant cell-sorting technologies with multiparameter selection capabilities. Here, we developed a cGMP-requirement-applicable fully closed cell sorter that has a suction mechanism and multiparameter detection using two laser optical settings. Negative pressure generated by a change in the chamber volume at a sorting point allows the isolation of cells of interest with high viability and purity. Our study demonstrated that this microfluidic sorter enables the isolation of cells of interest at an effective rate of 7,000 sorts per second on average. A purity of 85.5% and 77.1% effective yield with 93.7% viability was obtained when applying a target population of 35.9% in total (lymphocyte+CD8+) at 15,000 events per second (2 × 10 cells/mL). The sorted gene-modified T cells maintain largely unaltered proliferation, antigen recognition, cytokine release, and cytotoxicity functionalities.
通过分析接受过过继性免疫细胞疗法治疗的患者，在起始材料和输注材料中发现了各种免疫细胞表型，它们是持续缓解的决定因素。为临床应用分离这些特定表型需要符合现行药品生产质量管理规范（cGMP）且具备多参数选择能力的细胞分选技术。在此，我们开发了一种适用于cGMP要求的全封闭细胞分选仪，其具有抽吸机制，并使用两种激光光学设置进行多参数检测。分选点处腔室体积变化产生的负压能够以高活力和高纯度分离出感兴趣的细胞。我们的研究表明，这种微流控分选仪平均每秒能够以7000次分选的有效速率分离出感兴趣的细胞。当以每秒15000个事件（2×10个细胞/毫升）分选占总体35.9%（淋巴细胞+CD8+）的目标群体时，获得了85.5%的纯度、77.1%的有效得率以及93.7%的活力。分选后的基因修饰T细胞在很大程度上保持了未改变的增殖、抗原识别、细胞因子释放和细胞毒性功能。
