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. 2018 Apr 12;8:5907. doi: 10.1038/s41598-018-24074-y

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Robust cardiomyocyte differentiation from RhiPSCs in fully chemically defined conditions. (a) Schematic of cardiomyocyte differentiation protocol from RhiPSCs. The differentiation stages and media were shown above and under the arrows and timeline, respectively. MEF-CM, MEF conditioned medium; CDBM, cardiac differentiation basal medium. (b) Immunofluorescence staining (top panels) and FACS analysis (bottom panels) for cardiac markers: cTnT, MYH, and α-ACTININ at day 10 of differentiation. Nuclei were stained by DAPI (blue). Scale bars, 50 µm. (c) Cardiac differentiation efficiency on different matrices (Matrigel, VTN-N, and Synthemax). Representative immunofluorescence staining for α-ACTININ and NKX2.5 (left panels) and flow cytometry analysis for cTnT (right panel) (data are mean ± SEM, n = 3). Nuclei were stained by DAPI. Scale bars, 50 µm. (d) Cardiac differentiation efficiency, assessed by cTnT expression via flow cytometry, of 8 unique RhiPSC lines on Synthemax-coated plates (data are mean ± SEM, n = 3).