Fig. 6

Edited clones give rise to large numbers of active NK cells and retain functional kill switch. (a) Overview of differentiation protocol to generate and expand NK-cells from iPSCs. (b) Schematic overview of cluster formation in cell culture dispersing into suspension as hematopoietic progenitor cells. (c) Brightfield images showing the transition from endothelial monolayer into suspension cells. (d-e) Flow cytometry quantification of CD56 expression (d) and HLA-I levels (e). (f) Summary of cytotoxicity assay measuring killing of K562 cells by NK cells at three different ratios. (g) Schematic representation of suicide assay of wild-type and engineered cells using flow cytometry. (h-i) Flow cytometry quantification of HLA-I expression in wild-type NK cells and edited NK cells mixed 1:1 before iCASP9 activation (h) and after activation and recovery period (i).(j) Brightfield images of wells containing wild-type or edited NK cells after suicide gene activation and recovery period. HSPC = hematopoietic stem/progenitor cells; HE hemogenic endothelium. Scale bar = 100 µm.