Cultivation of Rat C6 Gliomas and Human Fibroblasts

The cell culture of rat C6 gliomas was obtained from the Russian Collection of Cell Cultures, Institute of Cytology, Russian Academy of Sciences (Saint-Petersburg, Russia). C6 rat glioma cells were cultured at 2.5 × 10⁵/mL in Petri dishes (d = 35 mm, Nunc, Allerod, Denmark) or 96-well flat-bottomed plates (TPP, Geneva, Switzerland, 1 × 10⁴/well) in Dulbecco’s Modified Eagle Medium (DMEM, Sigma-Aldrich, Darmstadt, Germany) containing 10% fetal bovine serum (Sigma-Aldrich) and 10⁻⁴ g/mL gentamicin sulfate (Shandong Weifang Pharmaceutical Factory Co., Liaocheng, China) in a Heracell CO₂ incubator (Thermo Fisher, Saint-Petersburg, Russia) at 37 °C with 95% humidity and 5% CO₂ for 1–2 days [44 ]. Fibroblast cell culture was obtained from the cell bank of Pokrovskaya Hospital (Saint-Petersburg, Russia). Human fibroblasts were cultured at 96-well flat-bottomed plates (TPP, 2 × 10⁴/well) in DMEM containing 10% fetal bovine serum and penicillin streptomycin 10⁻⁴ g/mL in a CO₂ incubator at 37 °C, 95% humidity and 5% CO₂ for 2 days [45 (link)].