This study investigates the effect of the structured surface to the adhesion, proliferation, and alignment of endothelial cells (HUVECs). The chemical state of laser structured surface was also investigated in comparison with non-treated surface. A novel design for biomedical applications consisting of parallel chain-like structures was realised on stainless steel surface by laser micromachining. The structures were designed to employ surface topography in the presence of micron and sub-micron features and to avoid intensive surface modification that could compromise the mechanical properties of thin devices like cardiovascular stents. The results showed that the structured surfaces favor the adhesion, proliferation, and alignment of HUVECs. The proliferation and the alignment of HUVECs were pronounced when periodic distance between two consecutive chain-like structures was 25 µm. Moreover, there was no significant difference of chemical composition on the structured surface suggesting that the cell proliferation and alignment were mainly influenced by the surface topography.