Glioblastoma mouse model

The construction of the orthotopic glioblastoma mouse model was referred to the previous study [23]. Firstly, the mice transplanted with the control or Romo1-overexpressed bone marrow were anesthetized with 0.75% pentobarbital sodium (50mg per kg body weight) and performed with craniotomy. Each mouse was carefully injected with 1×10⁵ of suspension GL261 cells through bregma with micro pump (2.5μl/min). Then, the injection hole was clogged with bone wax, and the incision was carefully stitched. Each group included at least 5 mice. The orthotopic tumor size was monitored by MRI, and the survival condition of each mouse was recorded after surgery.

For the treatment with clodronate liposomes (Liposoma BV) in the glioblastoma mouse model, 100μl of clodronate liposomes per mouse were injected through the tail vein every 3 days from Day 7 post orthotopic injection of GL261 cells. For the treatment with PD-1 antibody in the glioblastoma mouse model, the PD-1 antibody (RMP1-14, Bio X Cell) was intraperitoneally injected at 5 mg/kg every 3 days from Day 7 post orthotopic injection of GL261 cells. All the experiments were approved and supervised by the Animal Welfare and Ethics Committee of Yancheng City No.1 People’s Hospital.