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. MSCs were isolated from mouse bone marrow and their ability to differentiate into osteocyte and adipocyte lineages and their surface markers were evaluated. Then، t he 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. 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. 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.