Objective: To explore in vitro culture method and growth characteristics of human great saphenous vein smooth muscle cells (VSMCs). Methods: VSMCs were primarily cultured and passaged by digestive and explant attached method, and then the cells were subcultured. The cultured cells were identified by morphological observation, immunofluorescence staining and Coomassie brilliant blue staining. The survival percentage, proliferation ability and migration ability of cells were determined by trypan blue staining, MTT assay, drawing growth curve and wound healing assay. Results: he primary cultured VSMCs grew out of the edge of tissue clumps after incubation for 5-7 d. The passage cells maintained "hill-valley" pattern of growth. Positive staining of α-actin in cell cytoplasm was revealed by immunofluorescence, and the cytoskeleton structure was observed as blue dense bundles with Coomassie brilliant blue staining. Cell survival percentage was 97%. The growth curve of VSMC resembled “S” in shape, a significant change of optical density was detected after culture for 3–6 d, and the wound width of the serum-free cultured VSMCs changed markedly within the first 24 h. Conclusion: In vitro cultured VSMCs are characterized by contractile phenotype and high purity with good structure and function. Proliferation activity is higher after growth for 3–6 d and migration ability is strongest within the first 24 h of culture.