Identification of human vascular endothelial cells/rat vascular endothelial cells transfected with vitamin D receptor gene in vitro

Aim To Identify of human/rat vascular endothelial cells transfected with vitamin D receptor gene in vitro.Methods The plasmid expression vector with human pcDNATM6.2-GW/EmGFPmiR was amplified in E.coli.The plasmid was extracted,purified by EndoFree Plasmid Maxi Kit.The plasmid of pcDNATM6.2-GW/EmGFPmiR was analyzed and identified by enzymes digestion and sequencing analysis.human vascular endothelial cells /rat vascular endothelial cells was cultured and transfected VDR gene were selected with G418.The expression of bFGF mRNA and its products in the transfected BMSCs were detected by real time PCR,immunohistochemistry,immunofluorescence and Western-blot.The proliferative cycle of transfected BMSCs were examined by flow cytometry analysis.Results BMSCs expressing bFGF gene were obtained by transfection with pcDNA3.1-bFGF gene via Lipofectamine.The cell lineage was confirmed that the expressed position of bFGF was mainly in cytoplasm after transfection with bFGF gene.Higher proportion of cells in proliferation cycle was shown in transfected BMSCs compared with non-transfected BMSCs(P0.05).Concluson pcDNA3.1-bFGF can be transfected into BMSCs cultured in vitro via Lipofectamine.Genetic modification of BMSCs with bFGF may promote the proliferation of BMSCs.