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Characterization of iCell cardiomyocytes using single-cell RNA-sequencing methods

Characterization of iCell cardiomyocytes using single-cell RNA-sequencing methods

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SCHMID, Christina, Christian T. WOHNHAAS, Tobias HILDEBRANDT, Patrick BAUM, Georg RAST, 2020. Characterization of iCell cardiomyocytes using single-cell RNA-sequencing methods. In: Journal of Pharmacological and Toxicological Methods. Elsevier. 106, 106915. ISSN 1056-8719. eISSN 1873-488X. Available under: doi: 10.1016/j.vascn.2020.106915

@article{Schmid2020Chara-52185, title={Characterization of iCell cardiomyocytes using single-cell RNA-sequencing methods}, year={2020}, doi={10.1016/j.vascn.2020.106915}, volume={106}, issn={1056-8719}, journal={Journal of Pharmacological and Toxicological Methods}, author={Schmid, Christina and Wohnhaas, Christian T. and Hildebrandt, Tobias and Baum, Patrick and Rast, Georg}, note={Article Number: 106915} }

Schmid, Christina Attribution-NonCommercial-NoDerivatives 4.0 International Rast, Georg Hildebrandt, Tobias Introduction<br />Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes are being evaluated for their use in pharmacological and toxicological testing, particularly for electrophysiological side effects. However, little is known about the composition of the commercially available iCell cardiomyocyte (Fuijifilm Cellular Dynamics) cultures and the transcriptomic phenotype of individual cells.<br /><br />Methods<br />We characterized iCell cardiomyocytes (assumed to be a mixture of nodal-, atrial-, and ventricular-like cardiomyocytes together with potential residual non-myocytes) using bulk RNA-sequencing, followed by investigation of cellular heterogeneity using two different single-cell RNA-sequencing platforms.<br /><br />Results<br />Bulk RNA-sequencing identified key cardiac markers (TNNT2, MYL7) as well as fibroblast associated genes (P4HB, VIM), and cardiac ion channels in the iCell cardiomyocyte culture. High-resolution single cell RNA-sequencing demonstrated that both, cardiac and fibroblast-related genes were co-expressed throughout the cell population. This approach resolved two cell clusters within iCell cardiomyocytes. Interestingly, these clusters could not be associated with known cardiac subtypes. However, transcripts of ion channels potentially useful as functional markers for cardiac subtypes were below the detection limits of the single-cell approaches used. Instead, one cluster (10.8% of the cells) is defined by co-expression of cardiac and cell cycle-related genes (e.g. TOP2A). Incorporation of bromodeoxyuridine further confirmed the capability of iCell cardiomyocytes to enter cell cycle.<br /><br />Discussion<br />The co-expression of cardiac related genes with cell cycle or fibroblast related genes may be interpreted either as aberrant or as an immature feature. However, this excludes the presence of a non-cardiomyocyte sub-population and indicates that some cardiomyocytes themselves enter cell cycle. 2020 Baum, Patrick Rast, Georg Wohnhaas, Christian T. Baum, Patrick Wohnhaas, Christian T. eng Hildebrandt, Tobias 2020-12-21T09:39:50Z 2020-12-21T09:39:50Z Characterization of iCell cardiomyocytes using single-cell RNA-sequencing methods Schmid, Christina

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