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

Michael Rutten, L. B. Lester, D. L. Graham, J. Lee, Clifford 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)
JournalFASEB Journal
Volume11
Issue number3
StatePublished - 1997

Fingerprint

islets of Langerhans
Adenoviridae
transfection
Islets of Langerhans
Viruses
Transfection
Rats
apoptosis
Cytology
Insulin
Apoptosis
rats
Adenovirus E1A Proteins
Bromodeoxyuridine
insulin
Agar
Assays
Islets of Langerhans Transplantation
viruses
cell transplantation

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

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

Transfection of rat pancreatic islets with E1A-12S adenovirus prevents apoptosis. / Rutten, Michael; Lester, L. B.; Graham, D. L.; Lee, J.; Deveney, Clifford; Rabkin, J. M.

In: FASEB Journal, Vol. 11, No. 3, 1997.

Research output: Contribution to journalArticle

Rutten, M, Lester, LB, Graham, DL, Lee, J, Deveney, C & Rabkin, JM 1997, 'Transfection of rat pancreatic islets with E1A-12S adenovirus prevents apoptosis', FASEB Journal, vol. 11, no. 3.
Rutten, Michael ; Lester, L. B. ; Graham, D. L. ; Lee, J. ; Deveney, Clifford ; Rabkin, J. M. / Transfection of rat pancreatic islets with E1A-12S adenovirus prevents apoptosis. In: FASEB Journal. 1997 ; Vol. 11, No. 3.
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