Primary culture of endothelial cells from atherosclerotic human aorta

Protein phosphatase inhibitors and heat preconditioning prevent Hsp27 dephosphorylation, F‐actin disruption and deterioration of morphology in ATP‐depleted endothelial cells

“…EC were isolated from human aortas or umbilical veins and cultured according to described technique [26]. The confluent cell cultures of the 1 3 passages were taken for the experiments.…”

Distinct effects of heat shock and ATP depletion on distribution and isoform patterns of human Hsp27 in endothelial cells

“…EC derived from human aorta or umbilical vein were isolated and cultured as described [21]. Exponentially growing cell cultures of the first passage were used in all experiments.…”

Morphological alterations in endothelial cells from human aorta and umbilical vein induced by forskolin and phorbol 12-myristate 13-acetate: a synergistic action of adenylate cyclase and protein kinase C activators.

“…Primary cultures of human umbilical vein endothelial cells (HUVECs) and human aortic endothelial cells (HAECs) were obtained as described (15,16), with minor modificationdispase was used instead of collagenase. Seeding density was 5-8 X 104 cells per cm2.…”

“…Only confluent (7-9 days after seeding) cultures were used for experiments. The main characteristics of these cultures have been reported (16). For morphometry, cultures were stained with silver nitrate (17) and photographed.…”

“…Drugs were dissolved in dimethylsulfoxide or ethanol (Merck) and stored as stock solutions in aliquots at -80°C. Prior to use the aliquots were diluted with medium 199 and 0.5% BSA or with growth medium [medium 199 supplemented with 25 mM Hepes, 2 mM L-glutamine, 1 mM pyruvate, penicillin at 100 units/ml, streptomycin at 100 ,ug/ml, 10% (vol/vol) fetal calf serum] and sterilized by filtration through 0.22-,um filters (Nalgene).Primary cultures of human umbilical vein endothelial cells (HUVECs) and human aortic endothelial cells (HAECs) were obtained as described (15,16), with minor modificationdispase was used instead of collagenase. Seeding density was 5-8 X 104 cells per cm2.…”