Gibco astrocyte medium

The human astrocytes were purchased from Thermo Fisher Scientific (Waltham, USA) and were cultured in the GIBCO Astrocyte Medium (Thermo Fisher Scientific). The glioma cell lines including SHG44, U251, U87 and SHG139 were purchased from the ATCC (Manassas, USA), and the cells were cultured in the DMEM medium with 10% fetal bovine serum (FBS; Thermo Fisher Scientific). Cells were kept in the incubator supplied with 5% carbon dioxide at 37 °C. Human astrocyte cells were used as a normal control.

U251SP glioblastoma cells were obtained from the Memorial Sloan-Kettering Cancer Institute (New York, NY), and were grown in Dulbecco’s Modified Eagle Medium (Sigma-Aldrich, St. Louis, MO)^{30 (link)} supplemented with 10% fetal bovine serum and penicillin (100 U/ml)-streptomycin (100 μg/ml) in an atmosphere of 5% CO₂ at 37 °C. Human astrocytes (N7805100) were obtained from Thermo Fisher Scientific (Waltham, MA), and were grown in Gibco Astrocyte Medium in an atmosphere of 5% CO₂ at 37 °C.

The human glioma cell line, U251MG, was purchased from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). U-251MG cells were grown in Dulbecco’s modified Eagle’s medium (D-MEM; FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) supplemented with 10% fetal bovine serum (Biowest SAS, Nuaillé, France) and penicillin (100 units/mL) streptomycin (100 µg/mL) solution (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) in non-coated flasks. Cultures were maintained in a humidified atmosphere of 5% CO₂ and 95% air at 37 °C, and the medium was changed every 2–3 days.
Immortalized human normal astrocytes (HASTR/ci35) were kindly provided by Dr. Tomomi Furihata of Chiba University [36 (link)]. HASTR/ci35 cells were routinely grown at 33 °C with 5% CO₂/95% air in Gibco^® Astrocyte Medium (A1261301, Life Technologies, Carlsbad, CA, USA) supplemented with 1% N2 supplement, 10% fetal bovine serum (FBS), penicillin–streptomycin, and 4 µg/mL blasticidin S (SantaCruz Biotechnology, Inc., Dallas, TX, USA). For HASTR/ci35 cell differentiation, a culture medium without FBS was used, and the culture temperature was set at 37 °C.