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. 2018 Apr 26;8:6618. doi: 10.1038/s41598-018-24954-3

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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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96-well differentiation illustrates S1P/LPA-mediated enhancement of hiPSC-cardiomyocyte differentiation when added concurrently with Wnt activator CHIR99021. (A) Illustration of the ‘regular’ chemically-defined cardiac differentiation protocol utilized in this study. S1P/LPA was added at different time points during hiPSC-CM differentiation. (B) Representative 96-well immunofluorescence images for cardiac troponin T (TnT) in green and nuclear DNA in blue of 2D monolayer-based, chemically-defined differentiation of two poorly differentiating hiPSC lines into cardiomyocytes. Staining was performed in a 96-well plate format on day 8-post differentiation hiPSC-CMs. S1P, LPA, or both were added for days 0–2, 4–6 or 6–8 during the hiPSC-CM differentiation process. (C) Quantification of TnT positive cell numbers of total represented as percentages TnT positive cells for each time point when S1P, LPA or both were added. Error bars represent standard deviation. * indicates p < 0.05 versus control. Experiments were performed in 2 different hiPSC lines in 3–6 replicates.