Objective：To study an effective method for culturing of human vascular smooth muscle cells (hVSMCs) in tissue engineered blood vessels (TEBVs) in vitro.
Methods：A 3-cm segment of human long saphenous vein was harvested under sterile conditions. The primary culture and subculture were done by modified tissue-piece inoculation and trypsin digestion respectively. The cells were purified by mechanical treatment and differential attachment. PDGF was combined with high concentration of glucose DMEM as the culture medium. The cultured cells were identified by morphology and immunohistochemistry with contrast microscope and SABC kit，and RT-PCR method detected the expression of α-SMA and calponin 1.
Results：The cultured cells possessed“peak and valley”characteristics for VSMCs under microscope. Immunohistochemical staining showed positive expression of intracytoplasmic α-actin, and RT-PCR detected positve expression of α-SMA and calponin 1.
Conclusions：Culture medium of PDGF combined with high concentration of glucose DMEM,and with differential attachment method can provide highly purified hVSMCs with good structure and function.