HUVEC cells (Lonza, Basel; Switzerland) were cultured in Endothelial Cell Medium (Gibco/Thermo Fisher Scientific, Waltham, MA) supplemented with 5% heat-inactivated fetal bovine serum (FBS; Gibco/Thermo Fisher), 1% penicillin/streptomycin and vascular endothelial growth factor [VEGF] for rapid proliferation, as described [37 (link)]. The cells were cultured at 37°C in humidified 5% CO2, 95% air. The polyP preparations (“Na-polyP[Ca2+]”, “Ca-P particles” or “Ca-polyP-MP”) were added at a final concentration of 30 μg/mL.
After an incubation period for up to 3 d the cells were stained with the far-red fluorescent DNA dye Draq5 (Biostatus, Shepshed; UK) for nuclear visualization or with DAPI (#D9542; Sigma; 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride) for polyP detection [38 ].
After an incubation period for up to 3 d the cells were stained with the far-red fluorescent DNA dye Draq5 (Biostatus, Shepshed; UK) for nuclear visualization or with DAPI (#D9542; Sigma; 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride) for polyP detection [38 ].
Free full text: Click here
