Fig. 5: Functional evaluation of CAR-neutrophils loaded with R-SiO₂-TPZ nanoparticles using biomimetic glioblastoma (GBM) models in vitro.

a Schematic of our in vitro tumor model of GBM with blood-brain-barrier (BBB), which is composed of endothelial cells on the cell insert membrane and tumor cells in the bottom of the same transwell. b Transwell migration analysis of neutrophils at 12 h is shown. Anti-GBM cytotoxicity of indicated neutrophils at 24 h (c) and 36 h (d) was measured and quantified. e ELISA analysis of IL-6 and TNFα released from indicated neutrophils at 36 h was performed. f Second migration of different neutrophils at 48 h is shown. g Anti-GBM cytotoxicity of indicated neutrophils at 60 h was measured and quantified. h–j Schematic of neutrophil-infiltrated three-dimensional (3D) tumor model in vitro was shown in (h). i Representative fluorescent images of infiltrated neutrophils in the 3D tumor models were shown. DAPI was used to stain the cell nuclear and CD45 was used to stain neutrophils. Scale bars, 200 μm. Biological triplicates were performed independently. j The corresponding tumor-killing ability of indicated neutrophils was measured and quantified using cytotoxicity kit. Data are represented as mean ± SD of five independent biological replicates, one-way analysis of variance (ANOVA). Source data are provided as a Source Data file.