This microfluidic flow-stretch chip integrates fluid shear stress (FSS) and cyclic stretch (CS), two major mechanical stimulations in cardiovascular systems, for cultured cells. The model chip can deliver FSS and CS simultaneously or independently to vascular cells to mimic the haemodynamic microenvironment of blood vessels in vivo. By imposing FSS-only, CS-only, and FSS+CS stimulation on rat mesenchymal stem cells and human umbilical vein endothelial cells, we found the alignment of the cellular stress fibers varied with cell type and the type of stimulation. The flow-stretch chip is a reliable tool for simulating the haemodynamic microenvironment.