Primary cultures of HUVEC (kindly provided by Dr. Guttorm Haraldsen, Oslo University Hospital, Norway) were considered as passage 1 and aliquots were stored at −80 °C. HUVEC were grown in a supplemented MCDB 131 cell medium, designated “activation mix” (containing glutamax, hepes (all from Lonza, Basel, Austria), heat-inactivated FCS (7.5%, PAA, GE Healthcare, UK)), FGF2 (1 ng/mL, PeproTech Nordic, Stockholm, Sweden), human epidermal growth factor (EGF) (10 ng/mL, R&D Systems, Inc.), hydrocortisone (1 µg/mL, Sigma, St. Louis, MO, USA), 100 μg/mL penicillin and 100 μg/mL streptomycin) at 37 °C, 5% CO2 humidified incubator. For the proliferation assay, the control medium was supplemented with drugs or tumor samples according to the experimental condition tested. Confluent cell monolayers were exposed to trypsin (0.25%) for 2 min to detach the cells, whereupon the trypsin was neutralized with control medium. Detached cells were centrifuged at 1000 rpm for 5 min before used in experiments, or frozen down. HUVEC were used up to passage 6.

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