Fig. 2
From: Pluripotent stem cell-derived CAR-macrophage cells with antigen-dependent anti-cancer cell functions
CAR-iMac cells showed antigen-dependent phagocytosis and anticancer cell functions in vitro and in vivo. a Confocal microscopy pictures showing phagocytosis of K562 or K562-CD19 cells (red) by CAR (CD19)-iMac cells (green). b Flow cytometry showing phagocytosis of K562 or K562-CD19 cells by CAR (CD19)-iMac cells. c Western blotting showing phosphorylation of ERK and NF-κB P65 in CAR (CD19)-iMac cells in the indicated conditions. d qRT-PCR showing cytokine gene mRNA expression when CAR (CD19)-iMac cells were incubated with K562 or K562-CD19 cancer cells for 24 h. n = 3, error bar: standard error of the mean. e Top GO terms enriched in genes up-regulated in CAR-iMac cells. Right panel is GSEA analysis of “positive regulation of cytokine production.” f Top KEGG pathways enriched in genes up-regulated in CAR-iMac cells. Right panel is GSEA analysis of “antigen processing and presentation.” g Confocal microscopic images showing phagocytosis of OVCAR3 cells (red) by iMac or CAR (meso)-iMac cells (green). h Flow cytometry showing phagocytosis of OVCAR3 ovarian cancer cells by iMac or CAR (meso)-iMac cells. i GO term analysis with RNA-seq data showing the up-regulated genes in CAR (meso)-iMac cells. Right panel is GSEA analysis of “cytokine activity gene.” j qRT-PCR showing cytokine gene mRNA expression when iMac or CAR (meso)-iMac cells were incubated with OVCAR3 cells for 24 h. n = 3. Error bar: standard error of the mean. k 3 × 106 DiR dye-labeled iMac cells were intraperitoneally injected into NSG mice. n = 3. Error bars represent standard error of the mean. l 4 × 105 of luciferase-expressing ovarian cancer cells (HO8910) were intraperitoneally injected into NSG mice. Mice were treated 4 h later with I.P. injection of PBS, 4 × 106 iMac or 4 × 106 CAR (meso)-iMac cells. Bioluminescence showing tumor development on the indicated days. Statistical analysis was calculated via one-way ANOVA with multiple comparisons between the PBS group and the CAR-iMac group. *P < 0.05; **P < 0.01. m Quantification of tumor burden (total flux) by bioluminescent imaging on day 4, 11, 14, and day 25 after CAR-iMac treatment was plotted. Data are presented as the median ± SD, with statistical significance calculated via one-way ANOVA with multiple comparisons. *P < 0.05; **P < 0.01, ns not significant