[Comparison of different methods of pEGFP-actin plasmid transfection to the endothelium].
OBJECTIVE
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.


METHODS
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.


RESULTS
(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.


CONCLUSIONS
Gene transfection into ECV-304 with lipofection and electroporation methods exhibited better stability and repeatability.
Luo
Qin
Q
Institute of Burn Research, Southwestern Hospital, The Third Military Medical University, Chongqing 400038, P.R. China.
Qi
Jie
J
Chen
Jian
J
Luo
Xiangdong
X
Yang
Zongcheng
Z
D000199:Actins / Q000235:genetics; D002478:Cells, Cultured; D018274:Electroporation; D004730:Endothelium, Vascular / Q000502:physiology*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014162:Transfection / Q000379:methods*
D000199:Actins; D011993:Recombinant Fusion Proteins