Hair follicle stem cells (HFSCs) possess powerful expansion and multi-differentiation potential, properties that place them at the forefront of the field of tissue engineering and stem cell-based therapy. The aim of the present study was to investigate the differentiation of human HFSCs (hHFSCs) into cells of an endothelial lineage. hHFSCs were expanded to the second passage in vitro and then induced by the addition of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) to the culture medium. The expression levels of endothelial cell (EC)-related markers, including von Willebrand factor (vWF), vascular endothelial cadherin (VE)-cadherin and cluster of differentiation (CD)31, were detected by immunofluorescence staining, flow cytometric analysis and reverse transcription-polymerase chain reaction. The hHFSCs expressed vWF, VE-cadherin and CD31 when exposed to a differentiation medium, similar to the markers expressed by the human umbilical vein ECs. More significantly, differentiated cells were also able to take up low-density lipoprotein. The data of the present study demonstrated that an efficient strategy may be developed for differentiating hHFSCs into ECs by stimulation with VEGF and bFGF. Thus, hHFSCs represent a novel cell source for vascular tissue engineering and studies regarding the treatment of various forms of ischaemic vascular disease.