Frontiers in Immunology (Jul 2024)

Scalable process development of NK and CAR-NK expansion in a closed bioreactor

  • Xuening Wang,
  • Maeve Elizabeth Byrne,
  • Chang Liu,
  • Minh Tuyet Ma,
  • Dongfang Liu,
  • Dongfang Liu

DOI
https://doi.org/10.3389/fimmu.2024.1412378
Journal volume & issue
Vol. 15

Abstract

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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.

Keywords