Gentle macs dissociator and tumor tissue dissociation kit

Patient samples used were provided by the Biobank Core Facility at Barrow Neurological Institute. Samples were de-identified and conformed to the Biobank IRB protocol. Patient-derived cell lines (GB16 and GB71) were established from resected primary GBM tumor tissue. Tissue was processed using the Gentle MACS Dissociator and Tumor Tissue Dissociation kit (Miltenyi Biotec Inc.; Auburn, CA). Animal husbandry was performed according to the guidelines of St. Joseph Hospital and Medical Center and Barrow Neurological Institute under the Institutional Animal Care and Use Committee-approved protocol. Five- to six-week-old CrTac: NCr-Foxn1^nu nude mice (Taconic Biosciences; Hudson, NY) were used for in vivo orthotropic transplant of luciferized murine glioma model [44 (link)] (Ink4a/ARF^−/−; hEGFRvIII). For orthotopic transplants, 2 μL of dissociated cells at a density of 100,000 cells/μL were injected in the right striatum, as described previously [44 (link), 45 (link)]. In vivo tumor growth was measured by IVIS xenogen bioluminescence imaging (BLI) system after IP injection of 150 mg/kg Luciferin (Gold Biotechnology; St, Louis, MO) every week after 1-month post-surgery. Tumor-bearing animals were euthanized at the onset of neurological symptoms.