Tracking single hiPSC-derived cardiomyocyte contractile function using CONTRAX an efficient pipeline for traction force measurement

Gaspard Pardon; Alison S Vander Roest; Orlando Chirikian; Foster Birnbaum; Henry Lewis; Erica A Castillo; Robin Wilson; Aleksandra K Denisin; Cheavar A Blair; Colin Holbrook; Kassie Koleckar; Alex C Y Chang; Helen M Blau; Beth L Pruitt

doi:10.1038/s41467-024-49755-3

Tracking single hiPSC-derived cardiomyocyte contractile function using CONTRAX an efficient pipeline for traction force measurement

Nat Commun. 2024 Jun 26;15(1):5427. doi: 10.1038/s41467-024-49755-3.

Authors

Gaspard Pardon^{1

2

3

4

5}, Alison S Vander Roest^{1

3

6

7}, Orlando Chirikian⁸, Foster Birnbaum², Henry Lewis¹, Erica A Castillo^{1

4}, Robin Wilson¹, Aleksandra K Denisin¹, Cheavar A Blair^{1

4

9}, Colin Holbrook², Kassie Koleckar², Alex C Y Chang^{2

3

10

11}, Helen M Blau^{2

3}, Beth L Pruitt^{12

13

14

15}

Affiliations

¹ Departments of Mechanical Engineering and of Bioengineering, Stanford University, School of Engineering and School of Medicine, Stanford, CA, USA.
² Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA, USA.
³ Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.
⁴ Departments of Bioengineering and Mechanical Engineering, University of California, Santa Barbara, CA, USA.
⁵ School of Life Sciences, EPFL École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
⁶ Department of Pediatrics (Cardiology), Stanford University School of Medicine, Stanford, CA, USA.
⁷ Department of Biomedical Engineering, Michigan Engineering, University of Michigan Ann Arbor, MI, USA.
⁸ Biomolecular Science and Engineering Program, University of California, Santa Barbara, CA, USA.
⁹ Department of Physiology, University of Kentucky, Lexington, KY, USA.
¹⁰ Shanghai Institute of Precision Medicine and Department of Cardiology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200125, China.
¹¹ Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, CA, USA.
¹² Departments of Mechanical Engineering and of Bioengineering, Stanford University, School of Engineering and School of Medicine, Stanford, CA, USA. [email protected].
¹³ Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA. [email protected].
¹⁴ Departments of Bioengineering and Mechanical Engineering, University of California, Santa Barbara, CA, USA. [email protected].
¹⁵ Biomolecular Science and Engineering Program, University of California, Santa Barbara, CA, USA. [email protected].

Abstract

Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) are powerful in vitro models to study the mechanisms underlying cardiomyopathies and cardiotoxicity. Quantification of the contractile function in single hiPSC-CMs at high-throughput and over time is essential to disentangle how cellular mechanisms affect heart function. Here, we present CONTRAX, an open-access, versatile, and streamlined pipeline for quantitative tracking of the contractile dynamics of single hiPSC-CMs over time. Three software modules enable: parameter-based identification of single hiPSC-CMs; automated video acquisition of >200 cells/hour; and contractility measurements via traction force microscopy. We analyze >4,500 hiPSC-CMs over time in the same cells under orthogonal conditions of culture media and substrate stiffnesses; +/- drug treatment; +/- cardiac mutations. Using undirected clustering, we reveal converging maturation patterns, quantifiable drug response to Mavacamten and significant deficiencies in hiPSC-CMs with disease mutations. CONTRAX empowers researchers with a potent quantitative approach to develop cardiac therapies.

MeSH terms

Cell Differentiation / drug effects
Cells, Cultured
Humans
Induced Pluripotent Stem Cells* / cytology
Induced Pluripotent Stem Cells* / drug effects
Myocardial Contraction* / drug effects
Myocardial Contraction* / physiology
Myocytes, Cardiac* / cytology
Myocytes, Cardiac* / drug effects
Myocytes, Cardiac* / physiology
Single-Cell Analysis / methods
Software*

Abstract

MeSH terms

Grants and funding