Developing a porcine transplantation model: efficient gene transfer into porcine vascular cells.
BACKGROUND
The pig is generally regarded as likely to be the preferred donor animal in xenotransplantation. Although many hurdles remain to be cleared, it would be useful to be able to manipulate porcine endothelium genetically, among other reasons, to test approaches in the modulation of inflammation. However, as a nondividing cell, it is less easy to manipulate.


METHODS
The authors performed in vivo and in vitro gene transfection experiments using as an adjunct the DNA-binding agent 4',6'-diamidino-2-phenylindole (DAPI), which protects DNA from degradation.


RESULTS
The introduction of DAPI into a liposomal transfection system was able to increase in vitro transfection efficiency of both endothelial and vascular smooth muscle cells from the pig, even in the presence of small amounts of serum. This last observation encouraged the authors to use this system in vivo in porcine carotid arteries. In this model, the authors were also able to demonstrate a high degree of transfection efficiency using DAPI, which seemed to work by protecting DNA from degradation.


CONCLUSIONS
The authors believe this technique may allow them to address many biological questions relating to intervening in vascular disease, inflammation, and immune responses in the context of transplantation and beyond.
Chen
Daxin
D
Department of Immunology, Imperial College London, Hammersmith Hospital, London, United Kingdom.
Morgan
Finn
F
Berton
Ivan
I
Herbert
Paul E
PE
Lechler
Robert I
RI
Dorling
Anthony
A
Williams
Gordon
G
Warrens
Anthony N
AN
D000818:Animals; D001011:Aorta / Q000166:cytology; D002339:Carotid Arteries / Q000166:cytology; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000637:transplantation*; D005456:Fluorescent Dyes; D018014:Gene Transfer Techniques; D066298:In Vitro Techniques; D007211:Indoles; D013552:Swine; D014162:Transfection; D014183:Transplantation, Heterologous / Q000379:methods*
D005456:Fluorescent Dyes; D007211:Indoles; C007293:DAPI