Fig 5.

In vitro platforms used for screening engineered T cells. (A) In 3D lung (A549) and breast (MDA-MB-231) tumor models, cytotoxicity and migration of CAR T cells were examined. Nuclei are labeled with DAPI (blue) and control T cells and CAR T cells are labeled in green. CAR T cells showed enhanced migration into the tumor bulk compared to normal T cells. Scale bar is 100 µm. Adapted with permission from Wallstabe et al [212] (B) 3D microdevices were used to investigate cytotoxicity of engineered T cells under hypoxia. HepG2-Env target cells (red) were cultured with mRNA T cell receptor engineered T cells (green) overnight. T cells were more migratory in normoxia. Adapted with permission from Pavesi et al [213]. (C). A 3D tumor model was used to investigate CAR T cell killing efficacy under hypoxia. SKOV3 Human epithelial ovarian cancer cells were incubated with normal type (NT)-T cells or CAR-T cells at a 20:1 ratio for 24, 48, and 72 hours and labeled with viability dyes (live, green; dead, red). Scale bar is 1000 µm. Adapted with permission from Ando et al [214].