Prolonged gene expression in primary porcine pancreatic cells using an Epstein-Barr virus-based episomal vector.
Epstein-Barr virus (EBV)-based plasmids containing the origin of replication (oriP) and EBV nuclear antigen 1 (EBNA-1) are well known for the stable episomal maintenance in human cells. In order to clarify whether an EBV-based plasmid can be maintained stably in the porcine pancreatic cells which are the primary candidate sources of islet xenotransplantation, we constructed pEBVGFP encoding the green fluorescent protein (GFP). Monolayer culture of the porcine neonatal pancreatic cells was lipofected with pEBVGFP or pGFP which was derived from pEBVGFP by deleting out oriP and EBNA-1. pEBVGFP significantly prolonged GFP expression not only in human cell lines but also in the primary porcine pancreatic cells compared with pGFP. Interestingly, the duct cells that are believed as the pancreatic precursor cells were preferentially transfected and conveniently enriched among the mixed primary cell populations using a hygromycin B selection. To our knowledge, this is the first report suggesting the potential application of an EBV-based plasmid for the extended gene expression in the primary porcine pancreatic duct cells.
Min
Kyoung Ah
KA
Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, San 56-1, Shinlim-Dong, Kwanack-Ku, Seoul 151-742, Republic of Korea.
Oh
Sang Taek
ST
Yoon
Kun Ho
KH
Kim
Chong Kook
CK
Lee
Suk Kyeong
SK
D000818:Animals; D002478:Cells, Cultured; D002934:Cinnamates; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D006921:Hygromycin B / Q000031:analogs & derivatives* / Q000494:pharmacology; D007700:Kinetics; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D010179:Pancreas / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics*; D013552:Swine; D014162:Transfection; D014407:Tumor Cells, Cultured
D002934:Cinnamates; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D006921:Hygromycin B; C026273:hygromycin A