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Integrated Ca2+ flux and AFM force analysis in human iPSC-derived cardiomyocytes
- 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.
- Subjects :
- 0301 basic medicine
Atomic force microscopy
Clinical Biochemistry
chemistry.chemical_element
Calcium
Biochemistry
Contractility
03 medical and health sciences
Force analysis
030104 developmental biology
0302 clinical medicine
Ca2 flux
Background Correction
chemistry
Force dynamics
Biophysics
Induced pluripotent stem cell
Molecular Biology
030217 neurology & neurosurgery
Subjects
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