Standard eagle s medium

The evaluation of the toxicity of the IA-12 and IA-12/oNu systems was estimated by means of the multifunctional Cytell Cell Imaging system (GE Health Care Life Science, Sweden) using the Cell Viability Bio App. Cell lines M-HeLa (epithelial carcinoma of the cervix, HeLa subline, M-HeLa clone) were acquired from the Type Culture Collection of the Institute of Cytology (Russian Academy of Sciences, Saint Petersburg, Russia) and Chang liver normal cells from the N.F. Gamaleya Research Center of Epidemiology and Microbiology (Moscow, Russia). The cells were seeded in a concentration of 10⁵ cells/mL in 96-well plates (Eppendorf, Hamburg, Germany), then 150 μL of standard Eagle’s medium (PanEco, Moscow, Russia) was added per well, and incubation proceeded with CO₂ at 37 °C. The medium was then supplemented with 10% fetal calf serum and 1% nonessential amino acids. Twenty-four hours after seeding, the systems under study were added at preset dilutions, 150 μL to each well. The dilutions were prepared immediately in a nutrient medium. The experiments were repeated three times. Intact cells cultured in parallel with experimental cells were used as a control.

The M-HeLa clone 11 human, epithelioid cervical carcinoma, strain of HeLa, clone of M-Hela; human duodenal adenocarcinoma (HuTu 80); human breast adenocarcinoma cells (MCF-7) from the Institute of Cytology (Sankt-Petersburg) and human change liver from the collection and Research Institute of Virology, Russian Academy of Medical Sciences (Moscow) were used.
Cell viability was estimated using multifunctional Cytell Cell Imaging system (GE Health Care Life Science, Sweden) and Cell Viability Bio App. The cells were cultured in a standard Eagle’s nutrient medium (PanEco) and supplemented with 10% fetal calf serum and 1% nonessential amino acids. The cells were seeded in a concentration of 10⁵ cells/mL in 96-well plates (Eppendorf), then 150 μL of standard Eagle’s medium (PanEco, Russia) was added per well, and incubation proceeded with CO₂ at 37 °C. Twenty-four hours after seeding the cells into wells, the tested compounds were added at a preset dilution, 150 μL to each well. Each experiment was repeated three times. Intact cells cultured in parallel with experimental cells were used as a control. The calculation of the concentration of the drug causing inhibition of cell growth by 50% (IC50) was performed with an Internet tool: MLA—“Quest Graph™ IC50 Calculator” (AAT Bioquest, Inc., https://www.aatbio.com/tools/ic50-calculator, accessed on 25 July 2019).