Mouse diet

Adult random-bred albino mice of either sex weighing 22–28 g were included in the present study. The animals were obtained from the vivarium of the Central Scientific-Research Laboratory, the National University of Pharmacy, Kharkiv, Ukraine, and maintained at 19–24 °C and at 50% humidity in a well-ventilated room with a 12h light/dark cycle. Mice were housed in standard polypropylene cages with free access to food (“Mouse diet”, LabDiet, St. Louis, MO, USA) and water. However, food—not water—was withdrawn 12 h before and during the experiment. All the experimental protocols were approved by the Bioethics Commission of the National University of Pharmacy (protocol No. 2, February 17, 2016). Experiments were conducted in accordance with “Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes”.

Human cancer cell lines MDA-MB-468, DLD-1, and A431 that express EGFR and negative control (no EGFR expression) MDA-MB-435 cell line were obtained from ATCC (Rockville, MD, USA). Cells were propagated by serial passage in MEM/EBSS medium, supplemented with 10 % fetal bovine serum (Biochrom) at 37°C in a humidified atmosphere of 5 % CO2.
All animals used in imaging experiments were cared for and maintained under the supervision and guidelines of the University of Saskatchewan Animal Care Committee (protocol # 20160005). Female CD-1 nude mice were obtained from Charles River Canada (St-Constant, Quebec) at 4 weeks of age and housed in a 12 h light, 12 h dark cycle in a temperature and humidity controlled vivarium. Animals had libitum access to mouse diet (Lab Diet, St. Louis, Missouri) and water. After one week of acclimatization, mice were subcutaneously injected with a suspension of 5 – 10 × 10⁶ MDA-MB-468, DLD-1, A431 or MDA-MB-435 cells in 100 μL of a 1:1 mixture of serum-free MEM/EBSS medium (HyClone Laboratories, Logan, Utah) and Matrigel matrix basement membrane (Discovery Laboware, Inc. Bedford, MA) at the right? hind limb of each mouse. Tumor growth was followed with caliper measurements.