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A dual-reporter system for specific tracing of pancreatic ss-cell lines that non-invasively measures viable in vivo islet cells

Authors
 Suk Kyoung Kim  ;  Joo Young Kim  ;  Young Suk Choi  ;  Mi-Kyung Lee  ;  Byung Wan Lee  ;  Hyun Chul Lee  ;  Eun Jig Lee 
Citation
 BIOTECHNOLOGY LETTERS, Vol.32(1) : 53-57, 2010 
Journal Title
 BIOTECHNOLOGY LETTERS 
ISSN
 0141-5492 
Issue Date
2010
MeSH
Adenoviridae/genetics ; Animals ; Cell Line ; Cell Line, Tumor ; Dependovirus/genetics ; Genetic Vectors/genetics ; Green Fluorescent Proteins/genetics ; Green Fluorescent Proteins/metabolism ; Humans ; Insulin-Secreting Cells/cytology* ; Insulin-Secreting Cells/metabolism ; Insulin-Secreting Cells/transplantation* ; Islets of Langerhans/cytology* ; Islets of Langerhans/metabolism ; Luciferases/genetics ; Luciferases/metabolism ; Mice ; Mice, Nude ; Rats
Keywords
Adenovirus/adenoassociate virus hybrid vector ; Dual reporter ; GFP ; Insulin-producing cell ; Luciferase
Abstract
Islet transplantation is a potential treatment for type 1 diabetes. Currently, islet graft survival is measured using invasive methods to determine blood glucose, insulin, and C-peptide levels, even though these variables have limited value. To trace beta-cell survival and functional status, we constructed an adenovirus/adenoassociate virus hybrid vector (Hyb-DR) carrying two reporter genes, luciferase and green fluorescent protein (GFP), linked by the internal ribosome entry site and driven by the rat insulin II promoter. Luciferase activity increased and positive GFP expression was observed in beta-cell lines (MIN6N8 and INS-1E) infected with Hyb-DR. Using an in vivo imaging instrument, the GFP signal was detected in the flanks of nude mice 2 weeks after transplanting Hyb-DR-infected MIN6 cells into the kidney capsule. Coinfection of Hyb-DR with plasmids carrying beta-cell-specific transcription factors also resulted in expression of luciferase and GFP in the non-beta-cell lines (HepG2, FL83B, and YGIC5). Thus, the dual reporter system provided quantitative and visual information about the functionality of the islet mass and activation of the insulin gene.
Full Text
http://link.springer.com/article/10.1007%2Fs10529-009-0113-3
DOI
10.1007/s10529-009-0113-3
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Joo Young(김주영)
Lee, Byung Wan(이병완) ORCID logo https://orcid.org/0000-0002-9899-4992
Lee, Eun Jig(이은직) ORCID logo https://orcid.org/0000-0002-9876-8370
Lee, Hyun Chul(이현철)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/100474
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