Diabetes Metab J.  2011 Feb;35(1):72-79. 10.4093/dmj.2011.35.1.72.

Repeated Gene Transfection Impairs the Engraftment of Transplanted Porcine Neonatal Pancreatic Cells

• Seo MK
• Sun CL
• Kim JW
• Yoon KH
• Lee SK

• KMID: 2281495
• DOI: http://doi.org/10.4093/dmj.2011.35.1.72

Abstract

BACKGROUND
Previously, we reported that neonatal porcine pancreatic cells transfected with hepatocyte growth factor (HGF) gene in an Epstein-Barr virus (EBV)-based plasmid (pEBVHGF) showed improved proliferation and differentiation compared to those of the control. In this study, we examined if pancreatic cells transfected repeatedly with pEBVHGF can be successfully grafted to control blood glucose in a diabetes mouse model.
METHODS
Neonatal porcine pancreatic cells were cultured as a monolayer and were transfected with pEBVHGF every other day for a total of three transfections. The transfected pancreatic cells were re-aggregated and transplanted into kidney capsules of diabetic nude mice or normal nude mice. Blood glucose level and body weight were measured every other day after transplantation. The engraftment of the transplanted cells and differentiation into beta cells were assessed using immunohistochemistry.
RESULTS
Re-aggregation of the pancreatic cells before transplantation improved engraftment of the cells and facilitated neovascularization of the graft. Right before transplantation, pancreatic cells that were transfected with pEBVHGF and then re-aggregated showed ductal cell marker expression. However, ductal cells disappeared and the cells underwent fibrosis in a diabetes mouse model two to five weeks after transplantation; these mice also did not show controlled blood glucose levels. Furthermore, pancreatic cells transplanted into nude mice with normal blood glucose showed poor graft survival regardless of the type of transfected plasmid (pCEP4, pHGF, or pEBVHGF).
CONCLUSION
For clinical application of transfected neonatal porcine pancreatic cells, further studies are required to develop methods of overcoming the damage for the cells caused by repeated transfection and to re-aggregate them into islet-like structures.

Keyword

Diabetes mellitus; Hepatocyte growth factor; Porcine neonatal pancreatic cells; Transfection

MeSH Terms

Animals
Blood Glucose
Body Weight
Capsules
Diabetes Mellitus
Fibrosis
Graft Survival
Hepatocyte Growth Factor
Herpesvirus 4, Human
Kidney
Mice
Mice, Nude
Plasmids
Transfection
Transplants
Blood Glucose
Capsules
Hepatocyte Growth Factor