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NK cells have attracted widespread attention as a promising tool for immunotherapy in recent years due to their key role in tumor monitoring and cancer cell killing. The adoptive transfer of cytolytic NK is an attractive immunotherapy method for the treatment of lymphoma and other malignant tumors. However, due to their short persistence in vivo and impaired effector functions, most clinical successes are negligible. Therefore, an important challenge in improving the clinical applicability of NK cells is to expand ex vivo NK cells that show increased functionality when infused in vivo. Unfortunately, in efforts to induce expansion of NK cells, most reports show moderate expansion and indicate that additional stimulation is required.
Creative Biolabs has developed a novel technology based on nicotinamide (NAM), which has been successfully used to expand hematopoietic stem cells from allogeneic bone marrow cord blood. Our optimized protocol allows NAM to adapt to adult donor NK cells and regulate the characteristics and functions of NK cells expanded ex vivo.
NAM is a form of vitamin B3 that can directly regulate redox-sensitive enzymes, cell stress, cell metabolism, mitochondrial function, and gene expression. Studies have shown that NAM successfully enhances the expansion of functional NK cells in feeder-free culture with IL-2 and IL-15 stimulation. In preclinical studies, NK demonstrated cytotoxicity as well as enhanced in vivo survival cultured with NAM. Therefore, NAM-based NK expansion technology is a promising tool for NK-based adoptive therapy.
NK cells can be obtained from PBMCs of HLA-haploidentical or mismatched related donors. Higher NK cell purity was achieved by depleting CD3+ T cells and subsequently selecting CD56+ cells using immuno-magnetic beads. Following enrichment, NK cells are expanded ex vivo in flasks by co-culturing with IL-15 and NAM. The expanded NK cells are evaluated every 2-3 days by FACS to assess their phenotype and viability. Cytotoxicity assays are also conducted to assess their cytotoxicity against target tumor cells.
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