Type I interferon protects neurons from prions in in vivo models

Ishibashi, Daisuke; Homma, Takujiro; Nakagaki, Takehiro; Fuse, Takayuki; Sano, Kazunori; Satoh, Katsuya; Mori, Tsuyoshi; Atarashi, Ryuichiro; Nishida, Noriyuki

To investigate the permeability of RO8191, the BBB Kit TM was used as an in vitro blood-brain barrier screening model (RBT-24 H, PharmaCo-Cell Co), consisting of co-cultures of endothelial cells, pericytes and astrocytes, according to the manufacturer's protocol (Pervin et al., 2017) . In brief, 10 mM of RO8191 was added to the luminal side as the blood-side in the BBB Kit TM using clear transwell inserts in 12-well plates. After incubation for 1 h, 200 ml of the luminal side medium and 900 ml of the abluminal side as the brain-side medium were harvested from the culture plate. The concentration of RO8191 (MW: 373.21) in each sample, including the rest of the administered original sample, were measured using lipid chromatography with tandem mass spectrometry (LC-MS/ MS). To quantify blood-brain barrier permeability, the apparent permeability coefficient (Papp) ( Â10 À6 cm/s) was calculated as described previously (Tominaga et al., 2015) .