Penicillin

Endothelial cells from human umbilical veins were isolated as previously described (Baudin et al., 2007 (link)). Umbilical cords were obtained from the perinatal center of the Almazov National Medical Research Center. Ethics committee of the Almazov National Medical Research center approved the research protocol for the study “Investigation of cellular and molecular bases of aortic pathologies using tissue obtained from the leftovers after surgical interventions”. Ethical permit number December 26, 2014. The form of uniformed content for the patients enrolled in the study is approved by Ethics committee of the Almazov National Medical Research center. Primary cultures of human umbilical vein endothelial cells (HUVEC) were grown in Endothelial Cell Basal Medium-2 (ECBM-2) (Promocell, Germany) with Supplement Mix (Promocell, Germany), 10% fetal calf serum (FCS) (HyClone, United States), 4 mM glutamine, 50 µg/ml penicillin, 50 µg/ml streptomycin in tissue culture flasks (Sarstedt, Germany) which had been pre-coated with 0.2% gelatin (Sigma, United States) at 37°C in a humidified atmosphere with 5% CO₂. Subculturing was performed twice a week. The monolayer disintegration was caused by cells incubation in Trypsin-EDTA solution (Sigma, United States). Cells from three to five passage were used in experiments.

Colon cancer cells (Caco‐2, HT‐29) and fibroblasts (CRL2072) were used to study the biological activity of the extracts. Cells were maintained in Dulbecco's modified Eagle's Medium (DMEM) supplemented with 10% heat‐inactivated fetal bovine serum, 2 mM l‐glutamine, and penicillin (100 U/ml)/streptomycin (100 g/ml) in flasks (75 cm² surface area; Sarstedt, Spain) at 37°C under 40% humidity with a 5% CO₂ atmosphere.