Vegf lifefactors kit

All cell lines were purchased from ATCC (Manassas, VA). Human aortic smooth muscle cells (HASMCs) were cultured in complete DMEM F12 medium containing 10% FBS, 50 μg mL^-1 penicillin/streptomycin and 2 mM glutamine. HASMCs (~85% confluence) received treatment in serum-free medium. Human umbilical vein endothelial cells (HUVECs) were cultured in VascuLife basal medium containing VEGF lifeFactors Kit (Lifeline Cell Technology, Frederick, MD). THP-1 cells and peritoneal macrophages were cultured in complete RPMI1640 medium containing 10% FBS, 50 μg mL^-1 penicillin/streptomycin and 2 mM glutamine.

EA.hy926 cells (purchased from ATCC), a human umbilical vein cell line, were cultured in DMEM medium containing 10% fetal bovine serum (FBS), 50 µg/mL penicillin/streptomycin. Human umbilical vein endothelial cells (HUVECs) and human aorta endothelial cell (HAEC) were purchased from ATCC and cultured in Vasculife basal medium (EC medium) containing VEGF Lifefactors kit (complete EC medium, Lifeline Cell Technology, Frederick, MD, USA). Human aortic smooth muscle cells (HASMCs) were kindly provided by Dr. Xi-long Zheng from Calgary University in Canada and cultured in DMEM/F12 (V : V = 1 : 1) medium supplemented with 10% FBS, 50 µg/mL penicillin/streptomycin and 2 mM glutamine. HASMCs less than 8 passages were used for experiments. ECs received indicated treatment in serum-free medium.

Human brain microvascular endothelial cells (male, ACBRI 376) were obtained from Cell Systems (Kirkland, WA, USA) and cultured in Vasculife VEGF Basal Medium supplemented with the associated VEGF LifeFactors Kit (Lifeline Cell Technology, Frederick, MD, USA) with the following modifications. The LifeFactors Kit fetal bovine serum was not used and only 10 mL of the LifeFactors-provided L-glutamine was used as a supplement. In addition, iCell Endothelial Cell Medium Supplement (Cellular Dynamics International, Madison, WI, USA) was added to 10%. Cells were used between passages 3 and 6 for the initial toxicity studies and not past passage 8 for all other experiments.