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Attachment, replication and thrombogenicity of genetically modified endothelial cells
R A Kotnis1, M M Thompson, S L Eady
1Department of Surgery, University of Leicester, Leicester Royal Infirmary, U.K.
Summary
Genetically modifying human endothelial cells using electroporation maintains their normal function. These modified cells are suitable for seeding prosthetic vascular surfaces to improve graft patency.
Area of Science:
- Biomedical Engineering
- Vascular Biology
- Cellular Engineering
Background:
- Endothelial cell seeding enhances vascular graft patency by creating an anti-thrombogenic surface.
- Genetic modification of endothelial cells offers potential to further improve graft performance.
- Maintaining normal cell function post-modification is crucial for successful application.
Purpose of the Study:
- To assess the functional characteristics of endothelial cells after genetic modification via electroporation.
- To determine if electroporation affects endothelial cell replication, attachment, and prostacyclin release.
Main Methods:
- Human endothelial cells were electro-transfected with a test plasmid (pTCF).
- Transfection efficiency was determined using a single electric pulse (1000 V/cm, 12.8 ms).
- Functional assays compared transfected cells to control cells, measuring replication, attachment, and prostacyclin release.
Main Results:
- Electroporation achieved a 10% transfection efficiency.
- No significant differences were observed in replication rates between transfected and control cells (p = 0.76).
- Attachment (p = 0.43) and both basal (p = 0.89) and stimulated (p = 0.11) prostacyclin release were comparable between groups.
Conclusions:
- Electroporation is an effective method for genetically modifying endothelial cells.
- Genetically modified endothelial cells retain normal functional characteristics.
- These functionally normal, modified cells are viable for use in endothelial cell seeding of prosthetic vascular grafts.