Figure 2.

Effect of different media compositions on mono-cultures of human umbilical vein endothelial cells (HUVEC). The cells were cultured over 14 days in endothelial cell growth medium, consisting of basal medium and supplements (EGM complete), in a 1:1 mixture of hepatocyte culture medium and EGM complete (HCM-I + EGM complete) or in HCM enriched with endothelial cell growth supplements (HCM-I + EGM supplements). The graphs show time-courses of glucose consumption (A) and lactate production (B) as well as gene expression analyses of the endothelial cell markers platelet endothelial cell adhesion molecule 1 (PECAM1) (C) and von Willebrand factor (VWF) (D). Fold changes of mRNA expression were calculated relative to HUVEC before starting the experiments (d0) with normalization to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression by the ∆∆Ct method. The area under the curve was calculated for time-courses of biochemical parameters and differences between groups were detected using the unpaired, two-tailed Student’s t-test; for glucose consumption and lactate production n = 6, gene expression analysis n = 3, mean ± standard error of the mean.