Isolation and culture of human umbilical artery smooth muscle cells and gene transfection of TFPI gene

Abstract

Objective The aim of the present study is to investigate the effect of tissue factor pathway inhibitor （TFPI） gene on cell cycle of human vascular smooth muscle cells.Methods Human vascular smooth muscle cells were separated from human umbilical artery and identified by immunohistochemical staining.The cells were transfected with various amount of pIRES-TFPI plasmid （1,2,and 3 μg/ml,respectively）and the TFPI expression in the cells were analyzed by RT-PCR.MTT assay was employed to detect the effect of TFPI gene on the proliferation of human vascular smooth muscle cells.Results The proliferation of vascular smooth muscle cells was inhibited in pIRES-TFPI group 5 and 7 days after gene transfection when compared with that of pIRES 1-neo transfection group.Conclusion The overexpression of TFPI gene in human umbilical artery vascular smooth muscle cells may contribute to the suppression of the proliferation of cells by gene transfection. Key words: Human smooth muscle cell;  Tissue factor pathway inhibitor;  Restenosis