COMPARATIVE EVALUATION OF VIRAL AND NONVIRAL METHODS OF GENE DELIVERY TO MOUSE MESENCHYMAL STEM CELLS

Mesenchymal stem cells (MSCs) are attractive targets for cell and gene
therapy, because they can differentiate into many cell lineages. Hence, finding an efficient and suitable
method for transferring of genetic materials to these cells is very essential. In this study, we evaluated
the efficiency of two methods of gene transferring, viral and nonviral, in transfection of mouse MSCs,
comparatively.
Materials and methods: MSCs were isolated from mouse bone marrow and their ability to
differentiate into osteocyte and adipocyte lineages and their surface markers were evaluated. Then, the
efficiency of two methods of nonviral gene transferring; calcium phosphate, and cationic lipid reagents
(Lipofectamine™ LTX with Plus™ Reagent and TurbofectTM); and also lentivirus vector were
examined in transfection of mouse MSCs with green fluorescent protein (GFP) plasmid.
Results: The isolated MSCs successfully differentiated to osteocytes and adipocytes. They were
positive for CD24, CD29 and CD44; and negative for CD11b and CD45 cell surface markers.
Nonviral gene transferring methods were completely inefficient in transfection of mouse MSCs;
whereas calcium phosphate precipitate was completely toxic to mouse MSCs and cationic lipids only
transfected less than three percent of the cells. In contrast, high transfection rate and GFP expression
(above 70%) was seen with lentivirus vector.
Conclusions: it seems that mouse MSCs are refractory to nonviral gene transferring methods. In
contrast, lentivirus-mediated gene transfer methods may be an efficient tool for transfection of mouse
MSCs without any interference on their phenotype and differentiation potential