Figure 4.

ML NK cell from HD and CRC patient samples have improved response against CRC cells, including patient derived spheroids. a) HD cNK and ML NK cells were stimulated with patient derived spheroids (5:1 E:T ratio). b) Representative dot plot and c) summary data show IFN-γ production by cNK and ML NK cells. n = 7. d) annexin V/7-AAD flow cytometry-based killing assay on two patient-derived spheroid lines. Spheroids were partially dissociated and co-cultured with healthy donor cNK or ML NK cells at indicated E:T ratio for 4 hours and then processed into a single cells for flow cytometric analysis. Percent of specific killing was calculated as the decrement in annexin V/7-AAD negative viable tumor cells compared to tumor cells alone, n = 7–8. E), patient derived ML NK cells and cNK cells were incubated with HCT116 and HT29 cells (5:1 E:T ratio). f, Representative flow plot and g) summary data show the frequency of IFN-γ producing cells by patient cNK and ML NK cells in response to CRC cell lines. n = 8. Cytotoxicity of cNK and ML NK cells against GFP expressing h) HCT116 and I) HT29 was assessed using incucyte. Two-way repeated measures ANOVA (d). Two-way ANOVA with Sidak (c, g) and holm-Sidak posttest (h, i). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.