Gene transfer into the liver by plasmid injection into the portal vein combined with electroporation.
BACKGROUND
We investigated the optimum conditions for gene transfer into the liver by injection of plasmid via the portal vein combined with electroporation.


METHODS
In anesthetized 7-week-old male Shionogi Wistar rats, a tapered 3Fr. catheter was inserted through a branch of the colonic vein and its tip was secured into the portal vein. After clamping the vessels around the right and caudal lobes of the liver, FITC-oligodeoxynucleotide (ODN), green fluorescence protein (GFP) plasmid, or pCAGGS-luciferase was injected into the right and caudal lobes during electroporation using electric pulses applied to the caudal lobe only.


RESULTS
Transfected cells were identified by FITC-ODN and GFP fluorescence, and transfection efficacy was estimated by measurement of luciferase activity. FITC-ODN and GFP were detected in the caudal lobes of the liver. FITC-ODN was particularly present in hepatocytes surrounding the Glisson capsule and central veins, while GFP was detected in only hepatocytes surrounding the Glisson capsule. Luciferase activity in the liver was dependent on the plasmid concentration and voltage of the electric pulse, but independent of the volume of plasmid solution. Luciferase activity was limited in the right lobe, in which plasmid solution was injected but no electroporation was applied.


CONCLUSIONS
Our results showed local expression of the induced gene at the electroporation site, suggesting it is potentially useful for liver function support during recovery after extensive liver surgery and for the treatment of chronic and/or diffuse liver diseases.
Kobayashi
Shogo
S
Department of Surgery and Clinical Oncology, Graduate School of Medicine, Osaka University, Osaka, Japan.
Dono
Keizo
K
Tanaka
Toshiyuki
T
Takahara
Shiro
S
Isaka
Yoshitaka
Y
Imai
Enyu
E
Nagano
Hiroaki
H
Tomoaki
Kato
K
Umeshita
Koji
K
Nakamori
Shoji
S
Sakon
Masato
M
Monden
Morito
M
D000818:Animals; D018274:Electroporation; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D007275:Injections, Intravenous; D008099:Liver / Q000378:metabolism*; D008297:Male; D010957:Plasmids / Q000008:administration & dosage / Q000627:therapeutic use*; D011169:Portal Vein; D051381:Rats; D017208:Rats, Wistar
D005456:Fluorescent Dyes; D016650:Fluorescein-5-isothiocyanate