Establishing In Vivo Medulloblastoma Model

An in vivo medulloblastoma model was established as described before[58 (link)]. Briefly, 6–7 weeks old NOD-scid IL-2Rg null (NSG) mice (the Jackson Laboratory) were anesthetized with xylazine-ketamine, and right carotid artery was exposed. Dissociated Math1-Cre; ptch1-flox/flox cells (1×10⁶ cells in 4 μL HBSS)[58 (link)] were injected into the mouse brains using murine stereotaxic system (Stoelting Co). The coordinates were: 1.8mm to the right of bregma and 3mm deep from the dura. To disassociate the orthotopic tumor into the single cells for the serial implantation or the ex-vivo experiment, the tumor was dissected from the cerebellum, followed by digestion using Papain (10 U per mL), DNase (250 U) and L-cystein (2 mg) at 37°C for 30 min.