It has now been established that the 12S protein encoded by the E1A-adenovirus can activate quiescent cells to proliferate. The aim of the current study was to determine whether transfection with E1A-12S could extend the lifespan and functionality of pancreatic islets in culture. Rat pancreatic islets were isolated and transfected with adenovirus and a lipid carrier (Tfx, Promega). Positive clones were isolated using G418-resistance and an anti-E1A antibody (Oncogene Sci.). We found that the E1A-12S transfected islets had significantly increased proliferative rates as measured by MTT and BrdU assays, could maintain islet insulin granule morphology, and found they retained glucose-induced insulin responsiveness compared with cultures of nontransfected islets. The E1A-12S transfected islets also did not form foci in soft agar, and they had more immunofluorescence for the anti-apoptosis marker bcl-2 compared to nontransfected islets. Islets transfected with the control E1A mutant virus Ad5-dl312 were similar to nontransfected islets in their characteristics. Transfection with the E1A-13S virus produced extensive islet necrosis. Transfected islets will provide a useful model system for the study of islet cell biology and transplantation experiments.
|Original language||English (US)|
|State||Published - 1997|
ASJC Scopus subject areas
- Molecular Biology