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. 2014 Apr 25;3(2):e000693. doi: 10.1161/JAHA.113.000693

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© 2014 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley Blackwell.

This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

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Figure 1. — CsA promotes differentiation and expansion of cardiomyocytes from the mouse PSC‐derived Flk1⁺ MPCs in OP9 co‐culture system. A, Protocol for differentiation of cardiomyocytes in OP9 co‐culture system. B, Live cell image showing αMHC‐GFP⁺ cardiomyocytes in differentiating Flk1⁺ MPCs. C, Image showing cTnT⁺ sarcomeres and DAPI⁺ nuclei of differentiating Flk1⁺ MPCs. D, Relative total cell number incubated with indicated concentrations of CsA. Population of total cells without CsA was regarded as 100%. E, Percentage of cTnT⁺ cardiomyocyte population incubated with indicated concentrations of CsA. F, Phase‐contrast image showing beating cardiomyocyte foci (black arrow head) incubated with control vehicle (Con) and CsA (2 μg/mL). G, Comparison of the number of beating foci generated by control vehicle and CsA. H, Live cell images showing αMHC‐GFP⁺ cardiomyocytes. I, Percentage of αMHC‐GFP⁺ area in a live cell image. J, Images showing cTnT⁺ cardiomyocytes and DAPI⁺ nuclei. K, Comparison of cTnT⁺ area (%). L and M, Representative FACS analysis and the percentage of ESC‐derived cTnT⁺ cardiomyocytes. N and O, Representative FACS analysis and the percentage of iPSC‐derived cTnT⁺ cardiomyocytes. All scale bars, 100 μm. *P<0.01 vs 0 or Con. CsA indicates cyclosporin A; cTnT, cardiac Troponin T; DAPI, 4,6‐diamidino‐2‐phenylindole; ESC, embryonic stem cell; FACS, fluorescence‐activated cell sorting; GFP, green fluorescent protein; iPSC, induced pluripotent stem cell; LIF, leukemia inhibitory factor; MHC, myosin heavy chain; MPCs, mesodermal precursor cells; SSC, side scatter.