Seeding of intravascular stents with genetically engineered endothelial cells.
The use of intravascular stents may be limited by both local thrombosis and restenosis due to intimal proliferation. In an effort to provide solutions to these problems, we seeded stents with genetically engineered endothelial cells in vitro. Using retroviral-mediated gene transfer, we inserted the gene for either bacterial beta-galactosidase or human tissue-type plasminogen activator (t-PA) into cultured sheep endothelial cells. The endothelial cells were seeded onto stainless steel stents and grown until the stents were covered. Expression of intracellular beta-galactosidase and high level secretion of t-PA were demonstrated both before and after the transduced cells were seeded onto the stents. Eight stents were expanded by in vitro balloon inflation, with observation of the seeded endothelial layer both prior to and after expansion. Most of the endothelial cells remained on the stents after balloon inflation. We conclude that intravascular stents can be coated with a layer of genetically engineered endothelial cells that can be either specifically labeled or made to secrete high levels of a therapeutic protein. Much of the layer of genetically engineered cells remains after the expansion of the stent in vitro. In vivo implantation of stents coated with genetically engineered endothelial cells may allow 1) introduction of genetically engineered endothelial cells directly into the vascular wall and 2) improvement of stent function through localized delivery of anticoagulant, thrombolytic, or antiproliferative molecules.
Dichek
D A
DA
Laboratory of Molecular Hematology, National Institutes of Health, Bethesda, Maryland 20892.
Neville
R F
RF
Zwiebel
J A
JA
Freeman
S M
SM
Leon
M B
MB
Anderson
W F
WF
D000818:Animals; D001807:Blood Vessel Prosthesis; D002404:Catheterization; D004730:Endothelium, Vascular / Q000166:cytology*; D005818:Genetic Engineering; D066298:In Vitro Techniques; D012756:Sheep; D013193:Stainless Steel; D015607:Stents; D010959:Tissue Plasminogen Activator / Q000235:genetics; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D013193:Stainless Steel; D001616:beta-Galactosidase; D010959:Tissue Plasminogen Activator