Medium 199

Human prostate tumour cell lines PC-3, DU-145 and LNCaP were obtained from DSMZ (Braunschweig, Germany). Tumour cells were grown and subcultured in RPMI 1640 (Gibco/Invitrogen; Karlsruhe, Germany). The medium contained 10% foetal calf serum (FCS), 2% 2-(4-(2-Hydroxyethyl)-1-piperazinyl)-ethansulfonsäure HEPES-buffer (1 M, pH 7.4), 2% glutamine and 1% penicillin/streptomycin. Subcultures from passages 7–11 were selected for experimental use.
Human endothelial cells (HUVEC) were isolated from human umbilical veins and harvested by enzymatic treatment with chymotrypsin. HUVEC were grown in Medium 199 (M199; Biozol, Munich, Germany), supplemented with 10% FCS, 10% pooled human serum, 20 μg/ml endothelial cell growth factor (Boehringer, Mannheim, Germany), 0.1% heparin, 100 ng/ml gentamycin and 20 mM HEPES-buffer (pH 7.4). Subcultures from passages 2–6 were selected for experimental use.

Caki-1 and KTCTL-26 cells were purchased from LGC Promochem (Wesel, Germany) and A498 cells from Cell Lines Service (Heidelberg, Germany). Tumor cells were grown and subcultured in RPMI 1640 medium (Seromed, Berlin, Germany) supplemented with 10% fetal calf serum (FCS), 20 mM HEPES-buffer, 1% glutamax, and 1% penicillin/streptomycin (all: Gibco/Invitrogen; Karlsruhe, Germany) at 37°C in a humidified, 5% CO₂ incubator. Subcultures from passages 5–24 were employed.
Human umbilical vein endothelial cells (HUVEC) were harvested by enzymatic treatment with dispase (Gibco/Invitrogen) and cultured in Medium 199 (M199; Biozol, Munich, Germany), supplemented with 10% FCS, 10% pooled human serum, 20 μg/ml endothelial cell growth factor (Boehringer, Mannheim, Germany), 0.1% heparin, 100 ng/ml gentamycin, and 20 mM HEPES-buffer (pH 7.4). Subcultures from passages 2–6 were employed.