Culture Conditions for Brain Cell Lines

The human GBM U87MG cell line, human brain endothelial hCMEC/D3 cell line, N2a cell line, BV2 cell line, U251 cell line, U251TR cell line were purchased from the the American Type Culture Collection (ATCC). U87MG-Luc cell line was purchased from Shanghai Model Organisms Center (Shanghai, China). The above cell lines were cultured in DMEM medium supplemented with 10% (V/V) fetal bovine serum and 1% (V/V) penicillin and streptomycin. The X01 glioblastoma stem cells (GSCs) were kindly provided by Professor Jong Bae Park from the National Cancer Center of South Korea, the more details are in the previous works^{49 (link),61 (link)–64 (link)}. X01 cells were maintained in DME/F12 supplemented with epidermal growth factor (EGF, 10 ng ml⁻¹), basic fibroblast growth factor (bFGF, 10 ng ml⁻¹), B27, and 1% penicillin and streptomycin. These cells were cultured as a monolayer in a humidified atmosphere containing 5% CO₂ at 37 °C.