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McMahon, JM,Conroy, S,Lyons, M,Greiser, U,O'Shea, C,Strappe, P,Howard, L,Murphy, M,Barry, F,O'Brien, T
2006
February
Stem Cells And Development
Gene transfer into rat mesenchymal stem cells: A comparative study of viral and nonviral vectors
Published
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RECOMBINANT ADENOASSOCIATED VIRUS ARTERY OCCLUSION MODEL MARROW STROMAL CELLS HUMAN ADIPOSE-TISSUE BONE-MARROW IN-VITRO PROGENITOR CELLS ADENOVIRAL VECTORS LENTIVIRAL VECTORS TRABECULAR BONE
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Mesenchymal stem cells (MSCs) have been proposed for use in combinatorial gene and cell therapy protocols for the treatment of disease and promotion of repair. The efficacy of such a therapeutic approach depends on determination of which vectors give maximal transgene expression with minimal cell death. The study was carried out on bone-marrow derived rat MSCs, and a range of vectors was tested on the same stem cell preparation. Adenovirus, adeno-associated virus (AAV; serotypes 1, 2, 4, 5, and 6), lentivirus, and nonviral vectors were compared. Lentivirus proved to be most effective with transduction efficiencies of up to 95%, concurrent with low levels of cell toxicity. Adenovirus also proved effective, but a significant increase in cell death was seen with increasing viral titer. Rat MSCs remained refractory to transduction by all AAV serotypes, in contrast to rabbit MSCs tested at the same time. Lipofection of plasmid DNA gave moderate transfection levels but was also accompanied by cell death. Electroporative gene transfer proved ineffective at the parameters tested and resulted in high cell death. High and moderate levels of cell transduction using lentivirus vectors did not affect the ability of the cells to differentiate down the adipogenic pathway.
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