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Fig. 6 | BMC Cancer

Fig. 6

From: An advanced glioma cell invasion assay based on organotypic brain slice cultures

Fig. 6

Multiple applications of the ex vivo invasion assay. Representative confocal images, scale bars 100 μm, image quality was optimized by adjustment of brightness, contrast and gamma. a-c Brain slices of genetically modified mice as a tool to modulate the microenvironment. Human glioma cell line LN319 implanted in a wild-type and b knockout brain slices. The deletion of a specific cell surface protein in the microenvironment significantly inhibits tumor cell invasion as quantified in (c). Error bars represent 95% confidence interval, (A) n = 7; (B) n = 5; **p < 0.001, Welch’s t-test. d Co-implantation of different cell types. Reactive astrocytes (pseudocolored in green, DiI labeled) and the murine glioma cell line SMA560 (red, DiD labeled) were co-cultured in a ratio of 3:2 as heterotypic multicellular spheroids and implanted in a wild-type brain slice. e-i The ex vivo invasion assay as a tool to identify invasion modulating compounds. 500 SMA560 glioblastoma cells were seeded per well to induce spheroid growth. 18 h prior to implantation, spheroids or brain slices were treated with 1 μM jasplakinolide. 24 h after implantation, brain slices were fixed and imaged by confocal microscopy. In contrast to e the highly invasive control-treated SMA560, f the treatment of the tumor cells or g brain slices with jasplakinolide significantly reduced their ability to invade without inducing cell death as examined by trypan blue staining (i). h Quantification of invasion. Error bars show 95% confidence interval, e n = 12; f, g n = 7; i n = 3; **p < 0.001; ***p ≤ 0.0001; Welch’s t-test, p-values Bonferroni corrected

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