Seon Pyo Hong 1, Sukhyun Song 1, Seungjoo Lee 2, Hyeonju Jo 3, Hyoung Kyu Kim 3, Jin Han 3, Jae-Hyeong Park 4, Sung Woo Cho 5,
Abstract
Background: Pluripotent stem cell-derived cardiomyocytes (CMs) have become one of the most attractive cellular resources for cell-based therapy to rescue damaged cardiac tissue.
Aim: We investigated the regenerative potential of mouse embryonic stem cell (ESC)-derived platelet-derived growth factor receptor-α (DGFRα)+ cardiac lineage-committed cells (CLCs), which have a proliferative capacity but are in a morphologically and functionally immature state compared with differentiated CMs.
Methods: We induced mouse ESCs into PDGFRα+ CLCs and αMHC+ CMs using a combination of the small molecule cyclosporin A, the rho-associated coiled-coil kinase inhibitor Y27632, the antioxidant Trolox, and the ALK5 inhibitor EW7197. We implanted PDGFRα+ CLCs and differentiated αMHC+ CMs into a myocardial infarction (MI) murine model and performed functional analysis using transthoracic echocardiography (TTE) and histologic analysis.
Results: Compared with the untreated MI hearts, the anterior and septal regional wall motion and systolic functional parameters were notably and similarly improved in the MI hearts implanted with PDGFRα+ CLCs and αMHC+ CMs based on TTE. In histologic analysis, the untreated MI hearts contained a thinner ventricular wall than did the controls, while the ventricular walls of MI hearts implanted with PDGFRα+ CLCs and αMHC+ CMs were similarly thicker compared with that of the untreated MI hearts. Furthermore, implanted PDGFRα+ CLCs aligned and integrated with host CMs and were mostly differentiated into α-actinin+ CMs, and they did not convert into CD31+ endothelial cells or αSMA+ mural cells.
Conclusion: PDGFRα+ CLCs from mouse ESCs exhibiting proliferative capacity showed a regenerative effect in infarcted myocardium. Therefore, mouse ESC-derived PDGFRα+ CLCs may represent a potential cellular resource for cardiac regeneration.
Keywords: Cardiomyocyte; Embryonic stem cell; Myocardial infarction; Pluripotent stem cell; Regeneration.
Figure 4 Proliferation and survival of implanted platelet-derived growth factor receptor-α+ cardiac lineage-committed cells in the infarcted heart. A: Representative confocal image of Ki-67+ host cardiomyocytes in the peri-infarcted region 3 d after cell injection. Dotted-lined rectangular region is magnified in right. Scale bar, 50 μm; B: Quantifications of Ki-67+α-actinin+ cardiomyocytes per 104 nuclei in the peri-infarcted region 3 and 15 d after the implantation. Each group, n = 4. aP < 0.05 vs myocardial infarction (MI); C: Representative confocal images of revascularization within the infarcted areas 15 d after the implantation. Scale bars, 100 μm; D: Quantifications of capillary density (No. of CD31+ ECs/mm2) within the infarcted areas. Each group, n = 4. bP < 0.01 vs MI; cP < 0.01 vs MI+ platelet-derived growth factor receptor-α (PDGFRα)+ cardiac lineage-committed cells (CLCs); E: Representative confocal images of tdTomato+ PDGFRα+ CLCs and αMHC+ CMs 60 d after the implantation. Three independent experiments showed similar findings. Scale bars, 20 μm; F: Schematic diagram illustrating the regenerative potential of PDGFRα+ CLCs in the infarcted heart. CLCs: Cardiac lineage-committed cells; CMs: Cardiomyocytes; MI: Myocardial infarction.
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