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Integrated Ca2+ flux and AFM force analysis in human iPSC-derived cardiomyocytes

Authors :
Jayakumar Rajadas
Yuanyuan Dai
Nadezda Ignatyeva
Gerd Hasenfuss
Andrey V Malkovskiy
Antje D. Ebert
Source :
Biological Chemistry. 402:113-121
Publication Year :
2020
Publisher :
Walter de Gruyter GmbH, 2020.

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.

Details

ISSN :
14374315 and 14316730
Volume :
402
Database :
OpenAIRE
Journal :
Biological Chemistry
Accession number :
edsair.doi...........3fee583902611550f9e630baa23ea7b6
Full Text :
https://doi.org/10.1515/hsz-2020-0212