title: Induction of efficient differentiation and survival of porcine neonatal pancreatic cell clusters using an EBV-based plasmid expressing HGF.
abstract: Porcine neonatal pancreatic cell clusters (NPCCs) have been actively studied as a source of pancreatic stem cell transplantation for the treatment of diabetes. In this study, the hepatocyte growth factor (HGF) gene was cloned in an Epstein-Barr virus (EBV)-based plasmid vector (pEBVHGF) and the effects of the HGF expression on the survival and differentiation of NPCCs were analysed. For comparison, pHGF was constructed by deleting EBNA-1 and OriP from pEBVHGF. The expression of HGF, as measured by ELISA, lasted longer when pEBVHGF was used than when pHGF was used. C-Met phosphorylation co-related with the expression of HGF in the transfected NPCCs. Immunocytochemistry experiments showed that NPCCs showed a higher and longer expression of insulin when they were transfected with pEBVHGF than with pHGF. Moreover, a greater number of NPCCs survived for a longer period after they were transfected with pEBVHGF than when they were transfected with pHGF. Taken together, these results indicate that transfecting NPCCs with the HGF gene using an EBV-based plasmid is a more effective method of inducing differentiation to beta cells and enhancing survival than using a conventional plasmid. Therefore, it may be possible to use EBV-based plasmids to modify pancreatic stem cells for xenotransplantation.
authors.affiliation: Research Institute of Immunobiology, Department of Biomedical Sciences, Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Min Sun
authors.initials: MS
authors.lastname: Kim
authors.forename: Ji-Won
authors.initials: JW
authors.lastname: Kim
authors.forename: Chenglin
authors.initials: C
authors.lastname: Sun
authors.forename: Sang Taek
authors.initials: ST
authors.lastname: Oh
authors.forename: Kun Ho
authors.initials: KH
authors.lastname: Yoon
authors.forename: Suk Kyeong
authors.initials: SK
authors.lastname: Lee
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation; D002470:Cell Survival; D004797:Enzyme-Linked Immunosorbent Assay; D017228:Hepatocyte Growth Factor / Q000235:genetics*; D004854:Herpesvirus 4, Human / Q000235:genetics*; D007150:Immunohistochemistry; D010179:Pancreas / Q000166:cytology* / Q000378:metabolism; D010766:Phosphorylation; D010957:Plasmids; D013552:Swine; D014162:Transfection
chemical_list: D017228:Hepatocyte Growth Factor