Human NK Cell Enrichment and Expansion Using Defined GMP Reagents: Functional and Phenotypic Outcomes 2252995

Abstract Introduction Natural killer (NK) cells are a key component of the innate immune system and chimeric antigen receptor NK cells have shown promise in adoptive cell therapy. NK cell therapies require highly controlled expansion and deep characterization. Here we evaluated Cell-ViveTM GMP reagents for utility in NK cell isolation, activation, culture and characterization. Methods NK cells were isolated from leukopak mononuclear cells using two methods: (1) positive selection via sequential T cell depletion and NK cell enrichment with GMP anti-CD3 and anti-CD56 magnetic beads, and (2) negative selection using a GMP NK cell isolation kit. Following isolation, NK cells were activated using GMP anti-NKp46/CD2 magnetic beads and expanded 14 days in vitro under serum-free, xeno-free conditions supplemented with GMP-grade recombinant human IL-2 and IL-15. Results Expanded NK cells were assessed for viability, expansion, surface marker expression, cytokine production, and cytotoxicity. Both isolation strategies yielded highly viable NK cell populations, with different purity and expansion kinetic profiles. Functional analyses demonstrated effective cytokine secretion and target cell killing capacity post-expansion. Conclusion Our results provide comprehensive evaluation of NK cell enrichment and characterization using two different GMP-grade isolation kits. Further, we demonstrate the capability of the Cell-ViveTM GMP portfolio to support and develop sophisticated ex vivo processing workflows for NK cell-based therapies. Funding Source N/A Topic Categories Technological Innovations in Immunology (TECH)