Nmri nude female mice

The animal experiments were performed under a protocol approved by the Danish Animal Welfare Council, Ministry of Justice (2016-15-0201-00920). For the in vivo studies, U-87 MG.luc2 cells were cultured in Dulbecco’s modified eagle medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin (Thermo Fisher Scientific) at 37°C and in 5% CO₂. When cells reached ~70% confluence, they were harvested and 2–3 × 10⁶ cells in 100 μl of PBS were inoculated into the left flank of NMRI nude female mice (Janvier Labs, France). Animals were left to grow tumors until they reached ~300 mm³. From then on, tumors were measured every other day with the use of a caliper, and tumor volume calculated through the formula: (length × width²) × 0.5. When tumors reached ~1,000 mm³, animals were euthanized.

NMRI Nude female mice (Janvier labs) were orthotopically grafted directly in the cerebellum at 7 weeks with 3 × 10⁵ cells/5 μl of ICN‐MB‐PDX‐4 cells as described in Garancher et al (2018). After 3 days, mice were administrated 300 μl of LY2157299 (Galunisertib, AbMole Bioscience) orally at a dose of 75 mg/kg in 12% DMSO, 30% PEG, and water. Mice were treated 7 days a week twice a day until day 30. Mice were injected with Cisplatin (Sigma) in saline solution at a dose of 2 mg/kg intra‐peritoneally at days 4, 8, and 12 post‐grafting. Mice were euthanized when scientific and clinical end points were reached and brains were collected and fixed.

Experiments were performed on 8- to 12-week-old NMRI nude female mice from Janvier Labs (France). All mouse experiments were conducted according to local and international institutional guidelines and were approved by either the Animal Care Committee of the IRCAN and the regional (CIEPAL Cote d’Azur #187 and #188) and national (French Ministry of Research #03482.01/02482.2 and # 02973.01/02973.2) authorities.