Female swiss nu nu mice

Female swiss Nu/Nu mice were purchased from Charles River (Saint-Germain Nuelles, France). MMTV-PyMT (FVB/N strain) transgenic mice [26 (link),27 (link)] were bred and maintained within the accredited Mouse Facility and Transgenics GIGA platform of the University of Liège (Liège, Belgium), under pathogen-free conditions. All animal experiments were conducted in accordance with the Federation of European Laboratory Animal Science Associations (FELASA) and were approved by the local ethical committee of the University of Liège.

All experimental procedures were conducted in compliance with recommendations issued by the Comite d’Ethique Regional Languedoc-Roussillon - Comite d’Ethique pour l’Experimentation Animale and the Guidelines for the welfare of animals in experimental neoplasia. Female Swiss nu/nu mice (Charles River Laboratories) were housed under clean room conditions in sterile individually ventilated cages. Animals received sterile irradiated chow and water ad libitum.
Ten million SUM159 cells were inoculated subcutaneously in 150 μl of 50:50 medium/matrigel (BD biosciences) in the right hind flank region of 6-week old mice. When tumours reached approximately 100 mm³ animals were randomly assigned into five groups of 10 mice. Mice were then administered with (a) vehicle or (b) oral teriflunomide at a dose of 5 mg/kg once a day for 28 days, (c) 7.5 mg/kg PF477736 twice daily (6-hour interval) by i.p injections on days 2, 9, 16 and 23, (d) teriflunomide and PF477736 according to the same schedules as single regimens and (e) 20 mg/kg paclitaxel by i.p injection on days 1, 8 and 15. Animals were weighed twice a week and tumour volumes were calculated by caliper measurements once every three days using the following formula: V = (length × width × height × π/6). Animals were euthanised when tumour burden reached 1500 mm³.