Culturing and Differentiating Human Brain Cells

Human brain microvascular endothelial cells/conditionally immortalized clone 18 (HBMEC/ci18), human brain pericytes/conditionally immortalized clone 37 (HBPC/ci37), and human astrocytes/conditionally immortalized clone 35 (HASTR/ci35) were established by Prof. Furihata et al. [[11] (link), [12] (link), [13] (link), [14] (link), [15] ]. For maintenance, HBMEC/ci18 cultures were grown in VascuLife complete medium, and HASTR/ci35 and HBPC/ci37 cultures were grown in astrocyte growth medium and pericyte medium, respectively. All culture media contained 4 μg/ml blasticidin S. These cells were cultured at 33 °C for growth and at 37 °C for differentiation. Our specific experimental procedure has been standardized as the standard operating procedure (SOP) (Supplemental information).