Attachment, replication and thrombogenicity of genetically modified endothelial cells.
Endothelial cell seeding of prosthetic surfaces has been proposed as a technique to improve the patency of vascular grafts following arterial reconstruction. The introduction of specific recombinant DNA into seeded endothelial cells may enhance the anti-thrombogenic nature of the endothelial-blood interface with a consequent reduction in graft thrombosis. However, the successful use of genetically modified endothelial cells in the seeding process relies on the cells retaining normal function in terms of cellular replication, attachment and secretion of anti-thrombotic mediators. Successful genetic manipulation of human endothelial cells has been accomplished by viral and chemical methods.


OBJECTIVE
To study the functional characteristics of electrontransfected endothelial cells.


RESULTS
Endothelial cells were electro-transfected with the test plasmid pTCF at a transfection efficiency of 10% utilising a single electric pulse with an electric field of 1000 volts/cm and a time constant of 12.8ms. The functional status of transfected endothelial cells was then compared with a control endothelial cell population. There were no significant differences in replication (p = 0.76), attachment (p = 0.43), basal (p = 0.89) or stimulated (p = 0.11) prostacyclin release between transfected cells as compared with control endothelial cells.


CONCLUSIONS
Genetically modified cells are functionally normal, and may be used in endothelial cell seeding of prosthetic vascular surfaces.
Kotnis
R A
RA
Department of Surgery, University of Leicester, Leicester Royal Infirmary, U.K.
Thompson
M M
MM
Eady
S L
SL
Budd
J S
JS
James
R F
RF
Bell
P R
PR
D001807:Blood Vessel Prosthesis; D002448:Cell Adhesion; D002455:Cell Division; D017690:Cell Transplantation; D018274:Electroporation; D004730:Endothelium, Vascular / Q000166:cytology* / Q000502:physiology; D011464:Epoprostenol / Q000378:metabolism; D006083:Graft Occlusion, Vascular / Q000517:prevention & control*; D006801:Humans; D019868:Kanamycin Kinase; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D011138:Polytetrafluoroethylene; D011815:R Factors / Q000235:genetics; D013927:Thrombosis / Q000517:prevention & control*; D014162:Transfection / Q000379:methods; D014471:Umbilical Veins / Q000166:cytology
D011138:Polytetrafluoroethylene; D011464:Epoprostenol; D017853:Phosphotransferases (Alcohol Group Acceptor); D019868:Kanamycin Kinase