Nucl Med Mol Imaging.  2008 Oct;42(5):394-400.

Comparison of Human Sodium/Iodide Symporter (hNIS) Gene Expressions between Lentiviral and Adenoviral Vectors in Rat Mesenchymal Stem Cells

• Park SY
• Kim SJ
• Lee WW
• Lee H
• Kim HJ
• Chung JK
• Kim SE

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+/-4.7%, 54.0+/-6.4%, 85.7+/-8.7%, and 98.4+/-1.3% 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+/-6,659 picomole/106 cells) was greater than that in adeno-hNIS-rMSC at MOI 100 (6,168+/-2,134 picomole/10(6) 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.

Keyword

rat mesenchymal stem cell; lentivirus; adenovirus; human sodium iodide symporter

MeSH Terms

Adenoviridae
Animals
Blotting, Western
Clone Cells
Cloning, Organism
Gene Expression
Genes, Reporter
Homologous Recombination
Humans
Immunohistochemistry
Ion Transport
Lentivirus
Mesenchymal Stromal Cells
Rats
Sodium Iodide
Stem Cells
Symporters
Track and Field
Transgenes
Sodium Iodide
Symporters