Optimization of the transfection of human endothelial cells by electroporation.
A method for efficient transfection of human umbilical vein endothelial cells (HUVEC) is presented here. The procedure involves cell synchronization followed by electroporation and allows the detection of the activity of low-strength promoters. Sodium butyrate, added to the culture medium after electroporation, strongly potentiates the transcriptional activity of viral promoters. The method can be successfully applied to analyze the transcriptional activity of different promoters linked to reporter genes or to express foreign genes controlled by viral promoters.
Hôpital de Bicêtre, Le Kremlin-Bicêtre, France.
J L
JL
Schwachtgen
V
V
Ferreira
D
D
Meyer
D
D
Kerbiriou-Nabias
D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D002452:Cell Count; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D018274:Electroporation; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D010957:Plasmids; D011993:Recombinant Fusion Proteins; D014162:Transfection / Q000379:methods*; D014471:Umbilical Veins; D001616:beta-Galactosidase / Q000235:genetics; D014841:von Willebrand Factor / Q000032:analysis
D002087:Butyrates; D011993:Recombinant Fusion Proteins; D014841:von Willebrand Factor; D020148:Butyric Acid; D015500:Chloramphenicol O-Acetyltransferase; D001616:beta-Galactosidase