Fully closed cell sorter for immune cell therapy manufacturing

• Published in: Molecular therapy. Methods & clinical development 2023-09, Vol.30, p.367-376
• Main Authors: Matsumoto, Masahiro, Tashiro, Shinji, Ito, Tatsumi, Takahashi, Kazuya, Hashimoto, Gakuji, Kajihara, Junji, Miyahara, Yoshihiro, Shiku, Hiroshi, Katsumoto, Yoichi
• Format: Article
• Language: English
• Subjects: cell purification; cell sorter; cell sorting; cell therapy; cell therapy manufacturing; cGMP; flow cytometry; functional assay; Original
• ISSN: 2329-0501; 2329-0501
• DOI: 10.1016/j.omtm.2023.07.012

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 × 107 cells/mL). The sorted gene-modified T cells maintain largely unaltered proliferation, antigen recognition, cytokine release, and cytotoxicity functionalities. [Display omitted] Masahiro and colleagues developed a fully closed microfluidic chip cell sorter with a suction mechanism that can meet cGMP regulations and demonstrated its sorting performance. This sorter enables the isolation of cells using multiple parameters to provide high purity and high effective yield. The sorted cells maintained their functionality.