Comparison of different methods of pEGFP-actin plasmid transfection to the endothelium / 中华烧伤杂志

ABSTRACT

<p><b>OBJECTIVE</b>To screen the optimal transfection method for human umbilical vein endothelium, so as for us to establish a basic method for the further transfection with other objective genes.</p><p><b>METHODS</b>The pEGFP-Actin plasmid was employed as an ectogenetic gene and was transfected into ECV-304 endothelium cells by lipofection, DEAE-dextran or electroporation transfection methods, respectively. The cell death rate and transfection rate after the transfection were compared among the three methods.</p><p><b>RESULTS</b>(1) Twelve hours after the transfection, the cell death rate was 4% by all three methods, however, the cell death rate increased up to 50% at 60 hours after the transfection by DEAE-dextran. (2) The transfection rate of ECV-304 could reach 95% by all the three methods. But the fluorescent intensity was stronger in lipofection group compared with electroporation method.</p><p><b>CONCLUSION</b>Gene transfection into ECV-304 with lipofection and electroporation methods exhibited better stability and repeatability.</p>