Isolation and Culture of GBM-Derived Endothelial Cells

Surgical specimens from human patients with GBM ages 48 to 83 in all genders, multiple races, and ethnicities were collected at the Department of Neurosurgery of the Hospital of the University of Pennsylvania. The collection of human tissues in compliance with the tissue banking protocol was approved by the University of Pennsylvania Institutional Review Board, and written informed consent was obtained from each participant. ECs were isolated and verified as previously described (10 (link), 38 (link)). Tumor-derived single-cell suspensions were prepared by the tissue bank. Cell suspensions were subjected to magnetic-activated cell sorting (MACS) with anti-CD31 antibody-conjugated magnetic beads (Miltenyi Biotech, 130-091-935). All cells were used between passages 2 and 6. ECs were treated with harmine (10 to 40 μM; Sigma-Aldrich, 286044).