Dulbecco s modified eagle s medium

HUVEC were isolated de novo as previously described [24 (link),25 (link)] and cultured in EGM2 medium (Cambrex); peripheral blood monocytes were isolated from blood buffy coat by cold aggregation [26 ] and cultured in RPMI1640 and 10% fetal bovine serum (FBS); human foreskin fibroblasts were generated from skin biopsies and cultured in Dulbecco’s modified Eagle’s medium (DMEM) + 10% FBS, as previously described [27 (link)]; human astrocytes and hepatocytes were purchased from Lonza and cultured in AGM and HCM medium (Lonza), respectively. Bowes human melanoma cells, Huh7 and HepG2 human hepatoma cells and HT1080 fibrosarcoma cells were obtained from the DSMZ German Collection of Cell cultures and cultured in Dulbecco’s modified Eagle’s medium with 10% FCS; HeLa cervical cancer cells and NB4 acute promyelocytic leukemia cells were from the DSMZ and grown in RPMI, 10% FCS. The identity of all cell lines was authenticated by DNA profiling (performed at the DSMZ) and no rodent mitochondrial DNA sequences could be detected. None of the cells studied had sequence variants in the proximal t-PA gene promoter.

Colorectal adenocarcinoma CaCo-2 cells (ACC169, DSMZ, Braunschweig, Germany) were cultured under standard cell culture conditions in Dulbecco’s modified Eagle’s medium with high glucose (4.5 g/L), supplemented with 10% heat inactivated Foetal Bovine Serum (North American origin), 1% penicillin–streptomycin, 4 mM l-glutamine and 1% non-essential amino acids. All cell culture reagents were purchased from Thermo Fisher Scientific, Italy. The experiments were performed between passages 1–10 after defrosting of cells from liquid nitrogen storage.