Fig. 2

Phenotypic analysis of pNK cells before and after expansion and after cryopreservation. a-e. Surface expression of activating receptors (a), co-activating receptors (b), inhibitory receptors (c), chemokine receptors (d), cytolytic proteins, and functional markers (e) of pNK cells from 4 different donors was analyzed using flow cytometry before (D0) and after expansion (D14) and after cryopreservation (D14 cryo). CD107a expression were assessed after NK cells were co-cultured with K562 cells for 4 h at the E:T ratio of 1:1. Statistical analysis was performed using a paired t-test (*P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001). D0 was compared with D14 or D14 cryo. Furthermore, D14 was compared with D14 cryo. f. pNK cells before (D0) and after (D14) NK cell expansion and after cryopreservation (D14 cryo) were cultured with K562 at the indicated E(Effector):T(Target) ratio for 4 h. pNK cell cytotoxicity against K562 was measured using the carboxyfluorescein succinimidyl ester/7-aminoactinomycin D (CFSE/7-AAD) assay. The assay was performed twice with expanded pNK cells from four different donors. Data are represented as mean ± SD. Statistical analysis was performed using two-way ANOVA (*P < 0.05 and **P < 0.01)