<i>Background/Aims:</i> Local haemodynamic and stromal microenvironments may determine the phenotype of endothelial cells (EC) and regulate their inflammatory responses. <i>Methods:</i> We compared neutrophil recruitment by EC from human umbilical veins (HUVEC) or arteries (HUAEC) or from human coronary arteries (HCAEC) after ‘static’ culture or exposure to shear stress (2 Pa for 24 h) and treatment with tumour necrosis factor-α (TNF-α) or interleukin-1β (IL-1β). <i>Results:</i> Static cultures of each type of EC recruited flowing neutrophils efficiently after treatment with TNF-α or IL-1β; differences in culture media caused minor variations. After shear conditioning, the response of HUVEC to TNF-α (but not IL-1β) was much reduced, while the responses of HUAEC and HCAEC to both cytokines were reduced. However, swapping the culture media suggested that the differences in the shear response arose largely from medium constituents, particularly basic fibroblast growth factor. When gene expression profiles for HUVEC were examined immediately after isolation, after 5 days in static culture and after re-exposure to shear, variations in gene expression were only partially attributable to the effects of changes in shear stress. <i>Conclusions:</i> The behaviour of cultured EC may depend as much on the physico-chemical culture conditions as on their origins. The EC phenotype appears to be highly pliable, with environmental factors, such as shear stress and growth factors, modifying responses in an inter-linked manner.