title: Integrated Analysis of Contractile Kinetics, Force Generation, and Electrical Activity in Single Human Stem Cell-Derived Cardiomyocytes.
abstract: The quantitative analysis of cardiomyocyte function is essential for stem cell-based approaches for the in vitro study of human cardiac physiology and pathophysiology. We present a method to comprehensively assess the function of single human pluripotent stem cell-derived cardiomyocyte (hPSC-CMs) through simultaneous quantitative analysis of contraction kinetics, force generation, and electrical activity. We demonstrate that statistical analysis of movies of contracting hPSC-CMs can be used to quantify changes in cellular morphology over time and compute contractile kinetics. Using a biomechanical model that incorporates substrate stiffness, we calculate cardiomyocyte force generation at single-cell resolution and validate this approach with conventional traction force microscopy. The addition of fluorescent calcium indicators or membrane potential dyes allows the simultaneous analysis of contractility and calcium handling or action potential morphology. Accordingly, our approach has the potential for broad application in the study of cardiac disease, drug discovery, and cardiotoxicity screening.
authors.affiliation: Cardiovascular Research Center, Massachusetts General Hospital, Boston, MA 02114, USA; Harvard Medical School, Boston, MA 02115, USA; University Medical Center Groningen, University of Groningen, Groningen, 9713 GZ, the Netherlands.
authors.forename: Jan David
authors.initials: JD
authors.lastname: Kijlstra
authors.affiliation: Cardiovascular Research Center, Massachusetts General Hospital, Boston, MA 02114, USA; Harvard Medical School, Boston, MA 02115, USA; Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.
authors.forename: Dongjian
authors.initials: D
authors.lastname: Hu
authors.affiliation: Institute of Bioengineering and Nanotechnology, 138669 Singapore.
authors.forename: Nikhil
authors.initials: N
authors.lastname: Mittal
authors.affiliation: Department of Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Kausel
authors.affiliation: University Medical Center Groningen, University of Groningen, Groningen, 9713 GZ, the Netherlands.
authors.forename: Peter
authors.initials: P
authors.lastname: van der Meer
authors.affiliation: Reify Corporation, Saratoga, CA 95070, USA.
authors.forename: Arman
authors.initials: A
authors.lastname: Garakani
authors.affiliation: Cardiovascular Research Center, Massachusetts General Hospital, Boston, MA 02114, USA; Harvard Medical School, Boston, MA 02115, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA. Electronic address: domian@mgh.harvard.edu.
authors.forename: Ibrahim J
authors.initials: IJ
authors.lastname: Domian
mesh_terms: D000200:Action Potentials; D001696:Biomechanical Phenomena; D002118:Calcium / Q000032:analysis; D002454:Cell Differentiation; D002460:Cell Line; D006801:Humans; D007700:Kinetics; D009200:Myocardial Contraction; D032383:Myocytes, Cardiac / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology*; D059010:Single-Cell Analysis / Q000379:methods
chemical_list: D002118:Calcium