Nestin-Tv-a;Ink4a/Arf−/− mice (BL/6 background) and Nestin-Tv-a mice (BL/6 background) have been previously described and were bred within the Netherlands Cancer Institute (NKI) animal facility. The RCAS-PDGFB-driven mouse models of gliomagenesis (PDG) have been previously described16 ,26 (link),27 (link),70 (link)–72 (link). Briefly, glioblastomas were induced in 5–6-week-old male and female mice by intracranial injection of DF-1 cells expressing an RCAS vector encoding PDGF-B HA in Nestin-Tv-a;Ink4a/Arf−/− mice (PDG-Ink4a model), or DF-1 cells expressing PDGF-B HA and a short hairpin RNA targeting TP53 in Nestin-Tv-a mice (PDG-p53).
The PDG-Ink4a-OVA model was developed by cloning the OVA sequence into the RCASBP-Y vector. DF1 cells were transfected using the calcium phosphate transfection kit (ThermoFisher) to generate DF1-OVA cells. Successful transfection was confirmed by flow cytometry assessment of OVA expression (Extended Data Fig.2a ). To induce tumor development, Nestin-Tv-a;Ink4a/Arf-/- mice were intracranially injected with a 1:1 ratio of 200,000 DF1-PDGFB and 200,000 DF1-OVA cells. For the GL261 model, 20,000 GL261 cells were intracranially injected in C57BL/6JRj mice (Janvier labs) to induce tumor development. All animal studies were approved by the Institutional Animal Care and Use Committees of the NKI.
The PDG-Ink4a-OVA model was developed by cloning the OVA sequence into the RCASBP-Y vector. DF1 cells were transfected using the calcium phosphate transfection kit (ThermoFisher) to generate DF1-OVA cells. Successful transfection was confirmed by flow cytometry assessment of OVA expression (Extended Data Fig.
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