Transfection of rat pancreatic islets with E1A-12S adenovirus prevents apoptosis

M. J. Rutten, L. B. Lester, D. L. Graham, J. Lee, C. W. Deveney, J. M. Rabkin

Research output: Contribution to journalArticle

Abstract

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 languageEnglish (US)
Pages (from-to)A295
JournalFASEB Journal
Volume11
Issue number3
StatePublished - Dec 1 1997

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

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    Rutten, M. J., Lester, L. B., Graham, D. L., Lee, J., Deveney, C. W., & Rabkin, J. M. (1997). Transfection of rat pancreatic islets with E1A-12S adenovirus prevents apoptosis. FASEB Journal, 11(3), A295.