Sca-1+ Stem Cell Survival and Engraftment in the Infarcted Heart Dual Role for Preconditioning-Induced Connexin-43

Background-We report that elevated connexin-43 (Cx-43) in stem cells preconditioned with insulin-like growth factor-1 (IGF-1) is cytoprotective and reprograms the cells for cardiomyogenic differentiation. Methods and Results-Sca-1(+) cells were preconditioned with 100 nmol/L IGF-1 for 30 minutes followed by 8 hours of oxygen glucose deprivation to assess the cytoprotective effects of preconditioning. LDH release assay, cytochrome c release, and mitochondrial membrane potential assay showed improved survival of preconditioned Sca-1(+) cells under oxygen glucose deprivation compared with nonpreconditioned Sca-1(+) cells via PI3K/Akt-dependent caspase-3 downregulation. We observed PI3K/Akt-dependent upregulation of cardiac-specific markers including MEF-2c (2.5-fold), GATA4 (3.1-fold), and Cx-43 (3.5-fold). Cx-43 inhibition with specific RNA interference reduced cell survival under oxygen glucose deprivation and after transplantation. In vivo studies were performed in a female rat model of acute myocardial infarction (n=78). Animals were grouped to receive intramyocardially 70 mu L Dulbecco modified Eagles medium without cells (group 1) or containing male 1 x 10(6) nonpreconditioned Sca-1(+) cells (group 2) or preconditioned Sca-1(+) (group 3) cells labeled with PKH26. Survival of the preconditioned Sca-1(+) cells was 5.5-fold higher in group 3 compared with group 2 at 7 days after transplantation. Confocal imaging after actinin and Cx-43 specific immunostaining showed extensive engraftment and myogenic differentiation of preconditioned Sca-1(+) cells. Compared with group 2, group 3 showed increased blood vessel density (22.3 +/- 1.7 per microscopic field; P<0.0001) and attenuated infarction size (23.3 +/- 3.6%; P=0.002). Heart function indices including ejection fraction (56.2 +/- 3.5; P=0.029) and fractional shortening (24.3 +/- 2.1; P=0.03) were improved in group 3 compared with group 2. Conclusions-Preconditioning with IGF-1 reprograms Sca-1(+) for prosurvival signaling and cardiomyogenic differentiation with an important role for Cx-43 in this process. (Circulation. 2009; 119: 2587-2596.)