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Primary monolayer cultures were trypsinized, re-suspended in agarose gel medium : DMEM/F-12 containing 10% FBS, 100 units/ml penicillin G, 100 mg/ml streptomycin, and 50 mg/ml ascorbate, to a concentration of 210 4 cells/ml, then mixed with equal volume of 1% low-temperature melting agarose (Sigma-Aldrich, Steinheim, Germany) in agarose gel medium, giving a final concentration of 110 4 cells/ml suspended in 0.5% low-temperature melting agarose in agarose gel medium (suspension agarose)

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Karbach S, Jansen T, Horke S, Heeren T, Scholz A, Coldewey M, Karpi A, Hausding M, Kroller-Schon S, Oelze M, Munzel T, Daiber A (2012) Hyperglycemia and oxidative stress in cultured endothelial cellsa comparison of primary endothelial cells with an immortalized endothelial cell line