Abstract
We developed a new approach for combined analysis of calcium (Ca2+) handling and beating forces in contractile cardiomyocytes. We employed human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) from dilated cardiomyopathy (DCM) patients carrying an inherited mutation in the sarcomeric protein troponin T (TnT), and isogenic TnT-KO iPSC-CMs generated via CRISPR/Cas9 gene editing. In these cells, Ca2+ handling as well as beating forces and –rates using single-cell atomic force microscopy (AFM) were assessed. We report impaired Ca2+ handling and reduced contractile force in DCM iPSC-CMs compared to healthy WT controls. TnT-KO iPSC-CMs display no contractile force or Ca2+ transients but generate Ca2+ sparks. We apply our analysis strategy to Ca2+ traces and AFM deflection recordings to reveal maximum rising rate, decay time, and duration of contraction with a multi-step background correction. Our method provides adaptive computing of signal peaks for different Ca2+ flux or force levels in iPSC-CMs, as well as analysis of Ca2+ sparks. Moreover, we report long-term measurements of contractile force dynamics on human iPSC-CMs. This approach enables deeper and more accurate profiling of disease-specific differences in cardiomyocyte contraction profiles using patient-derived iPSC-CMs.
Funding source: Deutscher Akademischer Austauschdienst
Award Identifier / Grant number: 57381412 (N. Ignatyeva)
Funding source: Deutsche Forschungsgemeinschaft
Funding source: Deutscher Akademischer Austauschdienst
Funding source: Deutsches Zentrum für Herz-Kreislaufforschung
Funding source: Clinic for Cardiology and Pneumolgoy, Goettingen University, Medical Center
Award Identifier / Grant number: A. Ebert
Funding source: German Research Foundation (DFG)
Award Identifier / Grant number: SFB 1002 Project A12 (A. Ebert)
Acknowledgements
This work was supported by the Deutsche Forschungsgemeinschaft (German Research Foundation) Sonderforschungsbereich 1002, Projekt A12 (A.E.) as well as the German Academic Exchange Service (DAAD, N.I.). We are grateful for support by the Heidenreich von Siebold program at the University Medical Center Göttingen, as well as the DZHK (German Center for Cardiovascular Research), partner site Göttingen, Germany, and the Clinic for Cardiology and Pneumology at the University Medical Center, Göttingen University.
Author contribution: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Conflict of interest statement: The authors declare no competing interests.
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Supplementary Material
The online version of this article offers supplementary material (https://doi.org/10.1515/hsz-2020-0212).
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