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Murine melanoma cells B16F10 (American Type Culture Collection (ATCC), Manassas, VA, USA) were cultured in Advanced Minimum Assential Medium (AMEM; Thermo Fisher Scientific, Waltham, Massachusetts, USA), murine breast carcinoma cells 4T1 were cultured in Advanced RPMI 1640 (Thermo Fisher Scientific) in a humidified 5% CO2 atmosphere at 37 °C.
All media were supplemented with 5% fetal bovine serum (FBS, Thermo Fisher Scientific), 10 mM L-glutamine (Thermo Fisher Scientific), 50 mg/ml gentamicin (Krka, Novo mesto, Slovenia) and 100 U/ml penicillin (Sandoz, Holzkirchen, Germany).
Eight to ten-week old female C57BL/6NCrl and BALB/cAnNCrl (Charles River Laboratories, Wilmington, Massachusetts, USA) weighing 18–21 g were used in experiments. Mice were kept in quarantine for 2 weeks prior experiments. Throughout the quarantine and experiments mice were kept in a specific-pathogen-free housing conditions PE Gloves.
All procedures were performed in compliance with the guidelines for animal experiments of the EU directive (2010/63/EU) and the permission from the Veterinary Administration of the Ministry of Agriculture, Forestry and Food of the Republic of Slovenia (permission no. U34401-1/2015/26).
Permission was provided based on the approval of the National Ethics Committee for Experiments on Laboratory Animals Food gloves, which is under the auspices of Ministry of Agriculture, Forestry and Food of the Republic of Slovenia. In the experimental groups between 6 and 20 mice were included due to the high biological variability of tumor response to calcium electroporation Synthetic Gloves.