Objective:To establish the method of human vascular smooth muscle cells(hVSMCs)culture. Methods:Tissues were separated from segments of human adult aorta obtained at chest surgery. Small pieces of medial layer(about 1 mm3) were placed in flask and cultured in DMEM containing 20% heat-inactivated FCS and supplemented with sodium pyruvate(1 mmol/L),L-glutamine(1 mmol/L), penicillin(100 U/mL) and streptomycin(100 mg/mL). The cells were subcultured with 0.25% trypsin. Purity of cultures was assessed by positive immunostaining for -琢-smooth muscle actin(-琢-SMA). Results:Two weeks later,the colonies formed by migrated cells from the explants were collected at confluence and routinely subcultured. Cells used in the experiments were between passages 3 and 8. Immunofluorescence staining reavealed that VSMCs displayed elongated in shape with enormous a-SMA expression. Conclusion:The method of aorta explants culture can be used to get the VSMCs in vitro.