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• W2025809720 abstract "In vitro human embryonic stem cells (hESCs) and human inducedpluripotent stem cells (hiPSCs) are able to differentiate intofunctional cardiomyocytes (CMs). Traditional suspension methodfor 3D embryoid bodies (EBs) formation results in heterogeneousEB population. Here, we hypothesized that forming homogenous EBin size and shape allows studying the mechano-biologicalproperties of spontaneously beating clusters containing CMs.Using a defined number of single cells in AggreWell plate, weformed homogeneous EBs that can be efficiently differentiatedinto functional CMs by application of defined growth factors.We checked the expression of cardiac markers by qRT-PCR,immunocytochemistry and western blotting in both beating EBsand enzymatically-dissociated CMs and found increasedexpression of MYH6, MYH7 and RYR2 genes as well as sarcomericpattern for cardiac troponin T and alpha-actinin. Using atomicforce microscopy-based technique, we measured the beatingproperties of hESC-EBs and hiPSC-EBs and found a slower beatrate in hESC-CMs when compared to hiPSC-CMs (51 ± 5 vs 74 ± 7bpm) and similar contraction force (31 ± 7 vs 39 ± 9 nN). Bothcell types respond upon stimulation or inhibition ofbeta-adrenergic pathway and caffeine. Using Ca2+ imaging wealso demonstrated the functionality of RyR2 to release Ca2+from the sarcoplasmic reticulum and evaluated the contributionof IP3R. Thus, upon xestospongin C and histamine (IP3Rmodulators), spontaneous Ca2+ transients are maintainedconfirming the role of RyR2. Using the patch-clamp technique,we evaluated the cardiac population constituting the EBs andfound that more than 75% of excitable cells exhibit atrial-likeaction potentials. Our results indicated that themechano-biological properties of homogenous beating EBs can beinvestigated. They also indicated that spontaneous beating EBscontain mostly functional and organized atrial CMs." @default.
• W2025809720 created "2016-06-24" @default.
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• W2025809720 date "2014-01-01" @default.
• W2025809720 modified "2023-10-06" @default.
• W2025809720 title "Molecular and Functional Characterization of Uniform-Sized Beating Embryoid Bodies and Cardiomyocytes from Human Embryonic and Induced Pluripotent Stem Cells" @default.
• W2025809720 doi "https://doi.org/10.1016/j.bpj.2013.11.3135" @default.
• W2025809720 hasPublicationYear "2014" @default.
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