Cd1 nude

In vivo experiments were approved by the local veterinarian department and conducted in accordance with the Swiss law of animal protection. Female athymic nude mice (CD-1 nude), age 5–6 weeks, were obtained from Charles River Laboratories, Sulzfeld, Germany. U87MG cells and AR42J-cells were suspended in PBS (5 × 10⁶ cells in 100 μL) and subcutaneously inoculated on each shoulder. Two to three weeks later, when the tumor reached a size of about 300–500 mm³, the mice were used for the in vivo studies.

Animal procedures were in accordance with UK Animal law (Scientific Procedures Act 1986), including local ethics approval. Female, C57BL/6 wild‐type (6–8 weeks) and CD1‐nude (8–10 weeks) mice were purchased from Charles River laboratories (Kent, UK) and housed in a pathogen‐free facility with 12‐h light cycles. KPC cells were derived from KrasLSL^G12D/+;p53^R172H/+;Pdx1‐Cretg/+ (KPC) tumours. MC38 cells were purchased from American Type Tissue Collection (ATCC). Cell line authentication was performed using Short Tandem Repeat profiling (Cancer Research UK genomic facility, Leeds Institute of Molecular Medicine, March 2014). All cell lines were negative for mycoplasma (Lonza Mycoalert™ Test kit). MC38 (0.5 × 10⁶) or KPC (0.25 × 10⁶) cells were injected into the flank(s) of anaesthetised mice. Tumours were measured daily in three dimensions using digital callipers, and volume was calculated using the formula 0.5 × Length × Width × Height. When tumours reached 80 mm³, mice were randomly assigned to treatment groups. Anti‐CSF (Bioxcell, clone 5A1) was administered intraperitoneally at a dose of 10 mg/kg three times weekly, anti‐PD‐L1 (Bioxcell, clone 10F.9G2) at 10 mg/kg on days −1, 3, 6 and 9 and anti‐CD8a (Bioxcell, clone 2.43) at 250 μg on days −1, 3, 6 and 9. Radiation was initiated when tumours reached 100 mm³, delivered via a Gulmay 320 irradiator.

Female, 8- to 10-week-old CD1 nude were obtained from Charles River Laboratories (Wilmington, MA). All experiments were conducted with the approval of the University of Ottawa Animal Care and Veterinary Services. Intravenous (IV) administration of 100 μL containing 4e8 PFU of virus was performed through the tail vein. Direct intratumoral injections of virus were performed using a 50 μL volume, containing 4e8 PFU of virus, with a 27 gauge × 1/2′′ needle. Mice were monitored daily and euthanized at indicated time points or upon signs of morbidity by carbon dioxide narcosis. For the mouse tumor models, mice were either injected subcutaneously or intravenously with 1 e6 human Ewing sarcoma cells. Treatments began when subcutaneous tumors were visible and measure at least 13 mm³ or greater, whereas lung tumor bearing mice began treatments after 1 week of tumor implantation, a time determined from previous experiments. Tumor volumes in cubic millimeters were determined using digital calipers and the formulae (lager measurement/2) × (smaller measurement) [2 (link)]. End-points that dictated the sacrifice of mice included tumor volume >1800 mm³, skin ulcerations, weight loss <4.0 g, and hind limb paralysis. Three animals were used and each animal had bilateral tumors and the entire experiment was repeated.