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Figure 2 Transplantation of hESC-CMs in the Healthy Heart
(A) The transplanted area was localized by coinjection of 2-µm fluorescent beads (green), and the grafted cells were identified by immunostaining with antihuman mitochondria antibodies (red) (bar: 50 µm). (B) (Left) Hematoxilin and eosin staining depicting a cluster of grafted human embryonic stem cell-derived cardiomyocytes (hESC-CMs) (arrows). (Right) Immunostaining of the boxed area. In this example, enhanced green fluorescent protein-expressing hESC-CMs were used and were identified as yellow cells containing both troponin I (red) and enhanced green fluorescent protein (green) immunosignals. HC = human cardiomyocytes; R = rat (bar: 100 µm). (C) Immunostaining with antisarcomeric  -actinin (red) and antienhanced green fluorescent protein (green, right) antibodies. At this stage (36 h), the grafted hESC-CMs (arrows) displayed an immature, striated pattern (bar: 12 µm). (D) Assessment of the proliferation capacity of the hESC-CMs (36 h post-grafting) using antihuman Ki-67 (green) and antitroponin I (red) antibodies (bar: 10 µm). (E) (Left) Immunostaining of the grafted area (30 days post-transplantation) using antisarcomeric -actinin (red) and anti-Cx43 (white) antibodies. (Right) Superposition of the immunostaining results with antienhanced green fluorescent protein (green) antibodies. Note the relatively organized sarcomeric pattern (bar: 20 µm). (F) Immunostaining of the transplanted fluorescently labeled (yellow) hESC-CMs with anti-Ryanodine antibodies. Nuclei are counterstained with To-Pro3 (blue) in all immunofluorescent images.
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