Establishment and Maintenance of Medulloblastoma PDXs

Three Group 3 medulloblastoma xenografts^{52 (link),53 (link)} established from pediatric patients were used for experiments: D341 Med (D341), D384 Med (D384), and D425 Med (D425). These tumors were generously provided by Darell D. Bigner, MD, PhD, Duke Medical Center^{29 (link),30 (link)}. The xenografts were maintained in athymic nude mice (Envigo, Pratville, AL). After tumors were harvested, the cells were dissociated using a Tumor Dissociation Kit (Miltenyi Biotec, San Diego, CA) per manufacturer’s protocol. All cell lines were maintained in neurobasal medium (Life Technologies, Carlsbad, CA) supplemented with B-27 supplement without Vitamin A (Life Technologies), N2 supplement (Life Technologies), amphotericin B (250 μg/mL), gentamicin (50 μg/mL), L-glutamine (2 mM), epidermal growth factor (10 ng/mL; Miltenyi Biotec) and fibroblast growth factor (10 ng/mL; Miltenyi Biotec). The cells were kept under standard conditions at 37 °C and 5% CO₂. All three medulloblastoma PDXs were verified within the last 12 months using short tandem repeat analysis (Heflin Center for Genomic Sciences, UAB, Birmingham, AL).