Backgrounds: Adhesion is prerequisite for survival and also key factor for the differentiation of MSCs. To enhance cell adhesiveness on the extracellular matrix after transplantation into the infarcted myocardium, we genetically modified MSCs with transglutaminase II (tTG), co-receptor of integrin-associated cell adhesion, ex vivo before transplantation. Methods and Results: Modified MSCs with tTG showed 2.7-fold and over 2-fold higher of tTG expression and surface tTG activity, respectively, than those of control MSCs. Quantitative adhesion experiments demonstrated that overexpression of tTG increased adhesion of MSCs on fibronectin by 2-fold. Spreading and migration of tTG-MSCs were also increased by 2-fold and 1.5-fold, respectively. Remarkably, adhesion of tTG-MSCs on the cardiogel, cardiac fibroblast-derived 3D matrix, was increased by 4-fold. Down-regulation of tTG by tTG-specific siRNA molecules decreased markedly adhesion and spread of MSCs on fibronectin or cardiogel. tTG-MSCs displayed a significantly increase phosphorylation of focal adhesion related kinases, FAK, PI3K, and Src, in adhesive state on fibronectin compared to MSCs only. 1 week after induction of rat myocardial infarction using LAD ligation, MSCs labeled with 4���,6-diamino-2-phenylindole (DAPI) were implanted. After 4 days, survival of tTG-MSCs was 80% higher than that of control MSCs and in the former, migration of implanted cells was enhanced. After implantation of 2.0 X 105 of tTG transfected MSCs into infarcted myocardium, viable cells labeled with DAPI were identified in host myocardium in 4 weeks. The expression of cardiac troponin T (CTn T) and voltage-gated Ca2+ channel (CaV2.1) was increased. Conclusions: Our studies indicate that overexpression of tTG in MSCs enhance MSCs adhesiveness into a matrix as well as spreading and migration and also enhance the assembly of focal adhesion complexes in vitro, and finally viability after implantation. These data provide a novel therapeutic rational for the treatment of myocardial infarction and end-stage cardiac failure by genetic modification of MSCs with a therapeutic gene before transplantation