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. 2021 Jun 11;9:674260. doi: 10.3389/fbioe.2021.674260

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Copyright © 2021 Kahn-Krell, Pretorius, Ou, Fast, Litovsky, Berry, Liu and Zhang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Comparison of monolayer and suspension using the optimized protocol. (A) Flow cytometry staining for cTnT in suspension differentiated cardiomyoctes (red) compared with antibody isotype control (blue) with corresponding quantification. (B) Flow cytometry staining for iPSC markers (SOX2, SSEA4, and Tra-1–60) in monolayer and suspension (red) differentiated cardiomyoctes compared with iPSC (blue) positive control. (C) RT-qPCR analysis of (i) stem cell, (ii) mesoderm, and (iii) cardiac gene expression throughout differentiation for monolayer (solid) and suspension (dotted) techniques. (D) TEM images of (i) suspension and (ii) monolayer differentiated CMs with labeled characteristic features, N nucleus, GJ gap junction, ZL z-line, and MC mitochondria (Scale bar = 1 μm). Measurements of mean sarcomere length in each condition (n = 25). ^∗p < 0.0001.