Effects of glioblastoma cells and conditioned media on brain endothelial cells in vitro

Brain endothelial (bEnd.3) cells were used as a model of the BBB, co-cultured with glioblastoma (U87) cells or treated with conditioned media from U87 cells. The cell density of mono-cultured brain endothelial cells treated with glioblastoma conditioned media was evaluated through the sulforhodamine-B assay. The effect of U87 cells and conditioned media (CM) on the growth and migration of bEnd.3 cells was assessed using xCELLigence real-time cell analyzer and scratch assay, respectively. The barrier integrity of bEnd.3 cells were evaluated through the sodium fluorescein. The expression of the tight junction (TJ) proteins namely ZO-1, claudin-5, and occludin (presence or absence, size, and modification or degradation states) in brain endothelial cells treated with conditioned media and co-cultured with U87 cells were evaluated using the Western Blot assay.