Abstract
Pancreatic islets are notoriously difficult to efficiently transduce genes with viruses whether in vivo or ex vivo, the latter only transducing superficial layers of the islet. To improve efficiency of transduction, we explored surgical approaches to virus delivery in vivo. A technique was developed for retrograde surgical perfusion into the rat biliopancreatic duct with a test adenovirus containing a construct coexpressing green fluorescent protein, the latter for detection of infected cells. Pancreatic islets isolated after acute pancreatic infusion and cultured for 2 days showed expression in the entire islet and in almost all islets. When rats were recovered from the surgery, and then islets isolated at 1 and 8 weeks after surgery, we continued to see extensive islet green fluorescent protein expression, albeit at more reduced levels at 8 weeks. This strategy of surgical pancreatic ductal perfusion of viruses is an effective way to transduce or reduce gene expression in pancreatic islets for both acute and chronic study.
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