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Fig. 6. Intracoronary delivery of CSCs reduces infarct size by promoting myocardial regeneration. Evaluation of infarct dimension and myocardial repair. R, L, and F (Lower) correspond to the amount of myocardium remaining, lost, and formed, respectively. *P < 0.05 between untreated and treated rats. Data are mean ± SEM.

Fig. 7. Intracoronary delivery of CSCs and postinfarction ventricular remodeling. Illustrated are echocardiographic measurements of LV anatomical parameters in diastole. The injection of CSCs prevented the progressive adverse LV remodeling and improved the anatomy of the infarcted heart. *P < 0.05 between untreated and treated infarcted rats. Data are mean ± SEM.

Fig. 8. Effects of intracoronary delivery of CSCs on echocardiographic LV systolic function and anatomy. (A-D) Representative 2D (A, C) and M-mode (B, D) images from an untreated (A, B) and a treated (C, D) rat 35 days after coronary occlusion/reperfusion. Contractile activity is present in the treated animal (arrowheads) and absent in the untreated animal.

Fig. 9. Regenerating myocytes in the noninfarcted right ventricle. Transverse section of right ventricular myocardium in a treated rat at 35 days after infarction. Myocytes are identified by a -sarcomeric actin antibody staining (red). One myocyte (arrow) is positive for both EGFP and a -sarcomeric actin (yellow-green).

Fig. 10. Intracoronary delivery of CSCs and myocardial regeneration. Another example of regenerated infarcted myocardium is shown in panels A-C (arrows), first by a -sarcomeric actin staining (A, red), then by EGFP labeling (B, green) of a -sarcomeric actin^POS myocytes, and then by their combination (C, EGFP and a -sarcomeric actin; yellow-green). Scale bar = 100 m m.

Fig. 11. Expression of MEF2C in regenerated CSC-derived cardiomyocytes. (A) MEF2C alone (magenta); (B) MEF2C (magenta) and nuclei (blue) identified by DAPI; (C) MEF2C (magenta), EGFP (green), and nuclei (blue) identified by DAPI; (D) MEF2C (magenta), a -sarcomeric actin (red), and nuclei (blue) identified by DAPI. Scale bar = 10 m m.

Fig. 12. Intracoronary delivery of CSCs and myocardial regeneration. EGFP^POS (green) and troponin I^POS(red) small undifferentiated newly-formed myocytes within the infarcted region express in their plasma membrane N-cadherin (white). Scale bar = 10 m m.

Fig. 13. Intracoronary delivery of CSCs and myocyte regeneration. Distribution of myocyte classes according to their volume and presence (new cells) or absence (old cells) of EGFP in the cytoplasm.

Fig. 14. Intracoronary delivery of CSCs and generation of vascular structures. (A,B) newly-formed coronary arterioles within regenerated infarcted myocardium express EGFP and a -smooth muscle actin in smooth muscle cells (yellow-green fluorescence). (C) newly-formed capillary profiles (arrows) within regenerated infarcted myocardium express EGFP and von Willebrand factor in capillary endothelial cells (yellow-green fluorescence; arrows).

Fig. 15. Intracoronary delivery of CSCs and generation of vascular structures. (A) EGFP^POS (green), a -smooth muscle actin^POS (red) smooth muscle cells in resistance arterioles exhibit GATA-6 in the nucleus (white) and contain erythrocytes in their lumen (TER-119^POS, yellow). (B,C) Smooth muscle cells in regenerated coronary arterioles within the surviving myocardium (B) also express EGFP and a -smooth muscle actin (yellow-green) and endothelial cells in newly-formed capillaries (C) are positive for EGFP and von Willebrand factor (yellow-green, arrow). Scale bars = 10 m m.

Fig. 16. Intracoronary delivery of CSCs reduces infarct size by promoting myocardial regeneration in the absence of cell fusion. (A) Isolated preexisting (a -sarcomeric actin; red) and regenerated (EGFP^POS-a -sarcomeric actin^POS; yellow-green, arrows) myocytes. Scale bar = 50 m m. (B) Distribution of DNA content in non-cycling (red) and cycling (Ki67^POS; green) myocyte and lymphocyte nuclei.

Fig. 17. The border zone of the infarct in a CSC-treated heart shows an EGFP^POS myocyte (green) that expresses Ki67 in the nucleus (white, arrowhead). Scale bar = 10 m m.

Fig. 18. CSCs traverse the wall of coronary vessels migrating to the myocardium. (A) Detection by two-photon microscopy of numerous EGFP^POS clonogenic CSCs (green) located within the lumen of coronary vessels (rhodamine-labeled dextran; red) 20 min after cell injection. (B) At 12 h after reperfusion and 8 h after cell injection, EGFP^POS CSCs (green) are present throughout the ischemic myocardium. The leakage of dextran in the tissue (red) is apparent. Some residual vessels are still detectable. Two-photon microscopy: focal depth = 20 m m. Scale bars = 20 m m.

Fig. 19. CXCR4 expression in CSCs. Western immunoblot showing a 43 kDa band corresponding to CXCR4 in HL-60 and A-10 cell lines with high and low levels of CXCR4 expression, respectively. Strong immunoreactivity for CXCR4 was detected in protein extracts from EGFP^POS/c-kit^POS CSCs obtained from four different clones.