Scalable Process Development of NK and CAR-NK Expansion in a Closed Bioreactor Using 721.221-mIL21 Expansion System 2440

Abstract Description Production of large amounts of functional NK and CAR-NK cells represents one of the bottlenecks for NK-based immunotherapy. In this study, we developed a large-scale, reliable, and practicable NK and CAR-NK production using G-Rex 100M bioreactors, which depend on a gas-permeable membrane technology. This system holds large volumes of medium with enhanced oxygen delivery, creating conditions conducive to large-scale PBNK and CAR-NK expansions for cancer therapy. Both peripheral blood NK cells (PBNKs) and CAR-NKs expanded in these bioreactors retained similar immunophenotypes and exhibited comparable cytotoxicity towards hepatocellular carcinoma (HCC) cells akin to that of NK and CAR-NK cells expanded in G-Rex 6 well bioreactors. Importantly, cryopreservation minimally affected the cytotoxicity of NK cells expanded using the G-Rex 100M bioreactors, establishing a robust platform for scaled-up NK and CAR-NK cell production. This method is promising for the development of “off-the-shelf” NK cells, supporting the future clinical implementation of NK cell immunotherapy. Funding Sources This work was supported in part by HL125018 (DL), AI124769 (DL), AI129594 (DL), AI130197 (DL), and CA267368 (DL), Rutgers University-New Jersey Medical School Liu Laboratory Startup funding, New Jersey Commission on Cancer Research COCR24PRF004 (MTM), and 1S10OD026875-01A1 (ST). Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)