TGF-beta 2 treatment enhances cytoprotective factors released from embryonic stem cells and inhibits apoptosis in infarcted myocardium



D. K. Singla; R. D. Singla; S. Lamm;C. Glass


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Abbreviated Journal Title

Am. J. Physiol.-Heart Circul. Physiol.


iodoacetic acid; H9c2 cells; transforming growth factor-beta 2; myocardial infarction; BREAST EPITHELIAL-CELLS; GROWTH-FACTOR-I; OXIDATIVE STRESS; TISSUE; INHIBITOR; MOUSE HEART; DEATH; CARDIOMYOCYTES; PATHWAYS; PROTECTS; 3-KINASE; Cardiac & Cardiovascular Systems; Physiology; Peripheral Vascular; Disease


Singla DK, Singla RD, Lamm S, Glass C. TGF-beta 2 treatment enhances cytoprotective factors released from embryonic stem cells and inhibits apoptosis in infarcted myocardium. Am J Physiol Heart Circ Physiol 300: H1442-H1450, 2011. First published February 4, 2011; doi:10.1152/ajpheart.00917.2010.-We investigated whether factors released from mouse embryonic stem (ES) cells primed with and without transforming growth factor (TGF)-beta 2 inhibit iodoacetic acid (IAA)-and H2O2-induced apoptosis in the cell culture system as well as after transplantation in the infarcted heart. We generated conditioned media (CMs) from ES cells primed with and without TGF-beta 2 and determined their effects on IAA-and H2O2-induced apoptosis in H9c2 cells. We also transplanted both ES-CMs in the infarcted heart to determine the effects on apoptosis and cardiac function after myocardial infarction (MI) at day (D) 1 and D14. Terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) staining, apoptotic ELISA, and cell viability data demonstrated significantly (P < 0.05) reduced apoptosis with ES-CM compared with controls in both cell culture models. Moreover, TGF-beta 2-primed ES-CM (T-ES-CM) demonstrated enhanced beneficial effects, with further reduced (P < 0.05) apoptosis compared with ES-CM, suggesting the a presence of additional cytoprotective released factors after TGF-beta 2 treatment. Next, our in vivo apoptosis data suggested significant decrease in apoptosis with both ES-CMs compared with MI alone at D1 and D14. Notably, T-ES-CM demonstrated significant (P < 0.05) inhibition of apoptosis and fibrosis with improved cardiac function compared with ES-CM at D14, whereas no such effects were observed at D1. Next, we confirmed that apoptosis is mediated through a prosurvival Akt pathway. Moreover, we determined that after TGF-beta 2 treatment there was a two-to fivefold increase in cytoprotective released factors (interleukin-10, stem cell factor, tissue inhibitor of matrix metalloproteinase-1, and VEGF) with T-ES-CM compared with ES-CM. In conclusion, we suggest that factors released from ES cells with and without TGF-beta 2 treatment contain antiapoptotic factors that inhibit apoptosis in vitro and in vivo. We also suggest that T-ES-CM demonstrates additional beneficial effects that provide useful information for future therapeutic applications in regenerative medicine.

Journal Title

American Journal of Physiology-Heart and Circulatory Physiology





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