Recent advances have improved our understanding of natural killer (NK) cell-mediated alloreactivity after hematopoietic cell transplantation (HCT) or with adoptive transfer. NK cells contribute to a graft-versus-leukemia effect and may play a role in preventing graft-versus-host disease or controlling infectious diseases after allogeneic HCT. New discoveries in NK cell biology, including characterization of NK cell receptors and their interactions with self-HLA molecules and a better understanding of the mechanism of NK cell education have led to the development of novel strategies to exploit NK cell alloreactivity against tumors. While early studies using autologous NK cells lacked efficacy, the use of adoptively transferred NK cells to treat hematopoietic malignancies has been expanding. The production of allogeneic donor NK cells requires efficient removal of T- and B cells from clinical-scale leukapheresis collections. The goal of this chapter is to review NK cell biology, NK cell receptors, the use of NK cells as therapy and then to discuss the clinical decisions resulting in our current good manufacturing practices processing and activation of human NK cells for therapeutic use.