Ex vivo expansion of human NK cells using K562 engineered to express membrane bound IL21

Natural killer (NK) cells have gained signifi cant attention for adoptive immunotherapy of cancer due to their well-documented antitumor function. In order to evaluate the therapeutic effi cacy of NK cell adoptive immunotherapy in preclinical models with a potential for clinical translation, there is a need for a reliable platform for ex vivo expansion of NK cells. Numerous methods are reported in literature using cytokines and feeder cells to activate and expand human NK cells, and many of these methods are limited by low-fold expansion, cytokine dependency of expanded NK cells or expansion-related senescence. In this chapter, a robust NK cell expansion protocol is described using K562 cell line gene modifi ed to express membrane bound IL21 (K562 mb.IL21). We had previously demonstrated that this platform enables the highest fold expansion of NK cells reported in the literature to date (>47,000-folds in 21 days), and produces highly activated and pure NK cells without signs of senescence, as determined by telomere shortening.