Figure 2.

(A) Visualization of killing by CAR-NK-92 cells by fluorescence microscopy. A2780, SKOV3, and OVCAR3 cells were co-cultured with effector NK cells at an E/T ratio of 5:1. CD19NK cells were used as an antigen-specificity control. During the early process of death, the cells became smaller in size with condensed cytoplasm and tightly packed organelles. After cell shrinkage, massive membrane blebbing occurred, followed by separation of cell fragments into apoptotic bodies and subsequently loss of the GFP signal. (B) Fluoroskan results showing the killing effect of the CAR-NK-92 cells in ovarian cancer cell lines. GFP-expressing A2780, SKOV3, and OVCAR3 cells were seeded in flat-bottom 96-well plates at previously determined densities (A2780, 2 × 10⁴ cells/well; SKOV3 and OVCAR3 1.5 × 10⁴ cells/well). On the next day, NK-92 and CAR-NK-92 cells were added at the effector/target (E/T) ratio of 5:1. After removing culture medium containing NK cells and cell debris, residual attached cells were lysed with SDS, and fluorescence intensity was measured at excitation 485 nm/emission 520 nm using Fluoroskan Ascent™ FL. * indicates a significant difference calculated by two-way ANOVA, p < 0.05. Values represent the mean from two separate experiments each containing three samples.