Culturing Primary Rat Aortic Cells in Plasmax Media

Primary rat aortic endothelial cells (Cell Biologics, RN-6052) and rat aortic smooth muscle cells (Lonza, R-ASM-580) were cultured in physiologically representative media (Plasmax) formulated as described in [21] (link). Plasmax media was supplemented with 10% fetal bovine serum (Sigma, F1051), 1% penicillin/streptomycin (Sigma, P4333), 6.5 mM L-glutamine and 1 mM sodium pyruvate to ensure adequate growth of primary cells. Cells were incubated in physiologically relevant oxygen levels within a 5% CO₂/ 5% O₂ humidified incubator (Keeley and Mann 2019). Sub-culture was performed with 0.25% tryspin/EDTA solution (Sigma, T4049). Media was refreshed every 24 h. Cells were cultured for 7 days at a final ZnSO₄ concentration of 0 µM (Control), 5 µM, 50 µM.