Gene delivery to pancreatic exocrine cells in vivo and in vitro

Isabelle Houbracken, Luc Baeyens, Philippe Ravassard, Henry Heimberg

Onderzoeksoutput: Meeting abstract (Book)

Samenvatting

Gene delivery in pancreatic exocrine cells can be a useful tool for genetic lineage tracing, overexpression studies and RNAi-experiments. We have tested different transduction methods and viral vectors in vitro and in vivo in rat and mouse pancreas.
For in vitro transfection/transduction of rat exocrine cells we have used commercial lipofection reagents, adenoviruses overexpressing EGFP (Ad-CMV-EGFP), and Mokola and VSV-G pseudotyped lentiviruses overexpressing EGFP (Mok and VSV-G Le-CMV-EGFP). For in vivo transduction of mouse and rat pancreas we injected Ad-CMV-EGFP and VSV-G Le-CMV-EGFP intraparenchymally.
Lipofection of rat exocrine cell cultures is inefficient: maximally 3.7 % of total cells expressed EGFP on day 8 after transfection with pCMV-EGFP. We compared the efficiency of Mokola and VSV-G pseudotyped lentiviruses in transducing rat exocrine cells. Transduction of rat exocrine cells with VSV-G Le-CMV-EGFP with a multiplicity of infection of 50 results in 42 ± 2 % EGFP+ cells. The Mokola variant had a 10 times lower efficiency. Adenoviral transduction leads to even higher transduction efficiencies than the VSV-G lentiviruses but its usefulness for gene delivery in vitro is hampered by a drastic increase in cell death and disturbance of monolayer formation. On the other hand, we could not transduce pancreatic cells in vivo by intraparenchymal administration of lentiviruses in mouse and rat pancreas. However, in vivo administration of adenoviruses intraparenchymally in mouse and rat pancreas resulted in high transduction efficiencies. Most transduced cells are of acinar origin. Furthermore, also islet cells can be transduced in this way, albeit at a lower transduction efficiency.
Originele taal-2English
TitelPoster presentation Beta cell differentiation and regeneration workshop, Peebles, Scotland 2009
StatusPublished - 26 feb 2009
EvenementUnknown -
Duur: 26 feb 2009 → …

Conference

ConferenceUnknown
Periode26/02/09 → …

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