Sc1800

TMZ (T2577) and vincristine (V0400000) were from Sigma-Aldrich. Gemcitabine (S1714) was from SelleckChem. JAG-6A, CC-I, JAG-32, and JAG-79 were provided by Opal Oncology (Cambridge, UK). U87MG was obtained from the ATCC. KNS42 and SF188 cells were obtained from Professor Chris Jones (Institute of Cancer Research, London, UK). UP-029, SEBTA-003, SEBTA-023, and SEBTA-025 were cultured in DMEM (61965 Fisher Scientific) supplemented with 10% HIFBS (F7524-500ML Sigma-Alrich). SC1800 and CC2565 non-neoplastic astrocytes were purchased from Lonza and maintained in astrocyte growth medium supplemented with SingleQuots (CC-3187 Lonza) including (CC-4123) rhEGF, insulin, ascorbic acid, and L-glutamine. Cells were cultured under normoxic (21%) or hypoxic (1%) O₂.

Human cerebral microvascular endothelial cell line D3 (hCMEC/D3) cells, immortalized with hTERT catalytic subunits and simian vacuolating virus 40 large T antigens (26 (link)), were donated by Dr. Pierre-Olivier Couraud (Institut Cochin, INSERM, Paris, France). Human, healthy, cerebral cortex–derived astrocytes (line SC-1800) and human brain vascular pericytes (HBVPs) were purchased from Caltag Medsystems (Buckingham, United Kingdom). hCMEC/D3 cells were grown in endothelial basal medium 2 (Lonza, Basel Switzerland) supplemented with SingleQuots (Lonza) and 2% human serum (Sigma-Aldrich, St. Louis, MO, USA). SC-1800 were grown in astrocyte basal medium (AGM; Lonza) supplemented with SingleQuots (Lonza) and 3% human serum (Sigma-Aldrich). HBVP were cultured in pericyte basal medium (Caltag Medsystems), supplemented with pericyte growth factors (Caltag Medsystems) and 2% human serum (Sigma-Aldrich). All cells were grown in a 5% CO₂ atmosphere, 37°C incubator. Cell lines were authenticated as human with a microfluidic electrophoresis system incorporating a 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) to analyze short tandem repeat PCR fragments from 10 human genomic loci of human cell lines (27 (link)). Cells were routinely tested for mycoplasma with a kit from Lonza.