대한핵의학회지 (1967년~2009년)
Nucl Med Mol Imaging 2008;42(5)394~400
렌티바이러스와 아데노바이러스를 통하여 쥐의 중간엽줄기세포에 사람 나트륨/옥소 공동수송체 유전자를 전달하였을 때의 발현성능 비교
(Comparison of Human Sodium/Iodide Symporter (hNIS) Gene Expressions between Lentiviral and Adenoviral Vectors in Rat Mesenchymal Stem Cells)
Author 박소연1,*․김성진2,*․이원우1,3․이희란2․김현주1․정준기1,3․김상은1,3,
So Yeon Park1,*, Sung Jin Kim2,*, Won Woo Lee1,3, Heuiran Lee2, Hyun Joo Kim1, June-Key Chung1,3, and Sang Eun Kim1,3
Affiliation 서울대학교 의과대학 핵의학교실1, 의학연구원 방사선의학연구소3, 울산대학교 의과대학 미생물학교실2
1Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, Korea;2Department of Microbiology, University of Ulsan College of Medicine, Seoul, Korea;3Institute of Radiation Medicine, Medical Research Center, Seoul National Unive
Abstract

Purpose: Quantitative comparison of transgene expression within stem cells between lentivirus and adenovirus- mediated delivery systems has not been reported. Here, we evaluated the human sodium iodide symporter (hNIS) gene expression in rat mesenchymal stem cell (rMSC) transduced by lentivirus or adenovirus, and compared the hNIS expression quantitatively between the two delivery systems. Materials and Methods: Lentiviral-mediated hNIS expressing rMSC (lenti-hNIS-rMSC) was constructed by cloning hNIS gene into pLenti6/UbC/V5-DEST (Invitrogen) to obtain pLenti-hNIS, transducing rMSC with the pLenti-hNIS, and selecting with blasticidin for 3 weeks. Recombinant adenovirus expressing hNIS gene (Rad-hNIS) was produced by homologous recombination and transduction efficiency of Rad-hNIS into rMSC evaluated by Rad-GFP was 19.1¡¾4.7%, 54.0¡¾6.4%, 85.7¡¾8.7%, and 98.4¡¾1.3% at MOI 1, 5, 20, and 100, respectively. The hNIS expressions in lenti-hNIS-rMSC or adeno-hNIS-rMSC were assessed by immunocytochemistry, western blot, and I-125 uptake. Results: Immunocytochemistry and western blot analyses revealed that hNIS expressions in lenti-hNIS-rMSC were greater than those in adeno-hNIS-rMSC at MOI 20 but lower than at MOI 50. However in vitro I-125 uptake test demonstrated that iodide uptake in lenti-hNIS-rMSC (29,704¡¾6,659 picomole/106 cells) was greater than that in adeno-hNIS-rMSC at MOI 100 (6,168¡¾2,134 picomole/106 cells). Conclusion: Despite lower amount of expressed protein, hNIS function in rMSC was greater by lentivirus than by adenovirus mediated expression. Stem cell tracking using hNIS as a reporter gene should be conducted in consideration of relative vector efficiency for transgene expression. (Nucl Med Mol Imaging 2008;42(5):394-400)

Keyword rat mesenchymal stem cell, lentivirus, adenovirus, human sodium iodide symporter
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