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ORIGINAL RESEARCH article

Front. Immunol.
Sec. NK and Innate Lymphoid Cell Biology
Volume 15 - 2024 | doi: 10.3389/fimmu.2024.1412378

Scalable Process Development of NK and CAR-NK Expansion in a Closed Bioreactor

Provisionally accepted
Xuening Wang Xuening Wang 1Maeve E. Byrne Maeve E. Byrne 1Chang Liu Chang Liu 1Minh T. Ma Minh T. Ma 1Dongfang Liu Dongfang Liu 1,2*
  • 1 New Jersey Medical School, Rutgers, The State University of New Jersey, Newark, New Jersey, United States
  • 2 Rutgers Biomedical and Health Sciences, Rutgers, The State University of New Jersey, Newark, New Jersey, United States

The final, formatted version of the article will be published soon.

    Natural killer (NK) cells serve as the first line of defense that recognizes and kills tumor cells directly. NK cells can be genetically engineered with a chimeric antigen receptor (CAR) for the adoptive treatment of infections and cancers. 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 retainretained similar immunophenotypes and exhibitexhibited 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: NK cell, CD147-IL15-CAR-NK, expansion, HCC, bioreactor

    Received: 04 Apr 2024; Accepted: 04 Jul 2024.

    Copyright: © 2024 Wang, Byrne, Liu, Ma and Liu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Dongfang Liu, New Jersey Medical School, Rutgers, The State University of New Jersey, Newark, 07103, New Jersey, United States

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.