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Figure 3 Engraftment of Human Muscle-Derived Cells in the Infarcted Heart

(A) Masson's trichrome staining (muscle is stained red, collagen is stained blue) at the peri-infarcted area of the CD56⁺CD34⁺CD144⁺ cell-injected heart 2 weeks after cell transplantation. Scale bar equals 500 µm. The dotted line shows the engraftment area of human muscle-derived cells, (B) which corresponds to fast skeletal myosin heavy chain-positive (green) immunostaining. In the image, the fast skeletal myosin heavy chain-positive myofibers are stained green, nuclei are stained blue, and cardiac troponin I-positive cardiomyocytes are stained red. Only the engraftment regions expressed fast skeletal myosin heavy chain; normal myocardium was negative for fast skeletal myosin heavy chain. Scale bar = 50 µm. (C) CD56⁺CD34⁺CD144⁺ myoendothelial cells regenerated more fast skeletal myosin heavy chain-expressing myocytes than did CD56⁺CD34^–CD144^– myogenic and CD56^–CD34⁺CD144⁺ endothelial cells (*p < 0.05). (D) A few nuclei (blue) found in the fast skeletal myosin heavy chain-positive (green) engraftment area stained positive for human-specific proliferating cell nuclear antigen (red, arrows). Scale bar = 33.3 µm. (E to L) Expression of a cardiac cell markers by myoendothelial cells in vivo. (E) Engrafted myoendothelial cells expressing the nLacZ reporter gene (blue, arrowheads) colocalized with (F) cardiac troponin I (red, arrowheads) staining. 4',6-diamidino-2-phenylindole-only (G, blue, arrow) and colocalized with troponin I (H, pink and blue). (I) An nLacZ-expressing donor cell (blue, arrowhead) colocalized with the (J) cardiac troponin T marker (green, arrowhead). DAPI-only (K, blue, arrow) and colocalized with troponin T (L, green and blue). Scale bars = 50 µm.

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