title: Matrigel Mattress: A Method for the Generation of Single Contracting Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes.
abstract: BACKGROUND
  The lack of measurable single-cell contractility of human-induced pluripotent stem cell-derived cardiac myocytes (hiPSC-CMs) currently limits the utility of hiPSC-CMs for evaluating contractile performance for both basic research and drug discovery.
  
  
  OBJECTIVE
  To develop a culture method that rapidly generates contracting single hiPSC-CMs and allows quantification of cell shortening with standard equipment used for studying adult CMs.
  
  
  RESULTS
  Single hiPSC-CMs were cultured for 5 to 7 days on a 0.4- to 0.8-mm thick mattress of undiluted Matrigel (mattress hiPSC-CMs) and compared with hiPSC-CMs maintained on a control substrate (<0.1-mm thick 1:60 diluted Matrigel, control hiPSC-CMs). Compared with control hiPSC-CMs, mattress hiPSC-CMs had more rod-shape morphology and significantly increased sarcomere length. Contractile parameters of mattress hiPSC-CMs measured with video-based edge detection were comparable with those of freshly isolated adult rabbit ventricular CMs. Morphological and contractile properties of mattress hiPSC-CMs were consistent across cryopreserved hiPSC-CMs generated independently at another institution. Unlike control hiPSC-CMs, mattress hiPSC-CMs display robust contractile responses to positive inotropic agents, such as myofilament calcium sensitizers. Mattress hiPSC-CMs exhibit molecular changes that include increased expression of the maturation marker cardiac troponin I and significantly increased action potential upstroke velocity because of a 2-fold increase in sodium current (INa).
  
  
  CONCLUSIONS
  The Matrigel mattress method enables the rapid generation of robustly contracting hiPSC-CMs and enhances maturation. This new method allows quantification of contractile performance at the single-cell level, which should be valuable to disease modeling, drug discovery, and preclinical cardiotoxicity testing.
authors.affiliation: From the Departments of Pharmacology (T.K.F.), Cardiovascular Medicine (Y.W.C., J.E.H., C.C.L., C.C.H.), and Department of Medicine, Divisions of Cardiovascular Medicine and Clinical Pharmacology, Oates Institute for Experimental Therapeutics (L.W., B.C.K.), Department of Molecular Physiology and Biophysics (A.G.C.), Vanderbilt University School of Medicine, Nashville, TN; Departments of Cell and Developmental Biology (C.H.W.) and Biomedical Engineering (N.B., W.D.M.), Vanderbilt University, Nashville, TN; Research Medicine, Veterans Affairs TVHS, Nashville, TN (C.C.H.); and Stanford Cardiovascular Institute, Department of Medicine, Division of Cardiology and Department of Radiology, Stanford University School of Medicine, CA (J.C.W.).
authors.forename: Tromondae K
authors.initials: TK
authors.lastname: Feaster
authors.affiliation: From the Departments of Pharmacology (T.K.F.), Cardiovascular Medicine (Y.W.C., J.E.H., C.C.L., C.C.H.), and Department of Medicine, Divisions of Cardiovascular Medicine and Clinical Pharmacology, Oates Institute for Experimental Therapeutics (L.W., B.C.K.), Department of Molecular Physiology and Biophysics (A.G.C.), Vanderbilt University School of Medicine, Nashville, TN; Departments of Cell and Developmental Biology (C.H.W.) and Biomedical Engineering (N.B., W.D.M.), Vanderbilt University, Nashville, TN; Research Medicine, Veterans Affairs TVHS, Nashville, TN (C.C.H.); and Stanford Cardiovascular Institute, Department of Medicine, Division of Cardiology and Department of Radiology, Stanford University School of Medicine, CA (J.C.W.).
authors.forename: Adrian G
authors.initials: AG
authors.lastname: Cadar
authors.affiliation: From the Departments of Pharmacology (T.K.F.), Cardiovascular Medicine (Y.W.C., J.E.H., C.C.L., C.C.H.), and Department of Medicine, Divisions of Cardiovascular Medicine and Clinical Pharmacology, Oates Institute for Experimental Therapeutics (L.W., B.C.K.), Department of Molecular Physiology and Biophysics (A.G.C.), Vanderbilt University School of Medicine, Nashville, TN; Departments of Cell and Developmental Biology (C.H.W.) and Biomedical Engineering (N.B., W.D.M.), Vanderbilt University, Nashville, TN; Research Medicine, Veterans Affairs TVHS, Nashville, TN (C.C.H.); and Stanford Cardiovascular Institute, Department of Medicine, Division of Cardiology and Department of Radiology, Stanford University School of Medicine, CA (J.C.W.).
authors.forename: Lili
authors.initials: L
authors.lastname: Wang
authors.affiliation: From the Departments of Pharmacology (T.K.F.), Cardiovascular Medicine (Y.W.C., J.E.H., C.C.L., C.C.H.), and Department of Medicine, Divisions of Cardiovascular Medicine and Clinical Pharmacology, Oates Institute for Experimental Therapeutics (L.W., B.C.K.), Department of Molecular Physiology and Biophysics (A.G.C.), Vanderbilt University School of Medicine, Nashville, TN; Departments of Cell and Developmental Biology (C.H.W.) and Biomedical Engineering (N.B., W.D.M.), Vanderbilt University, Nashville, TN; Research Medicine, Veterans Affairs TVHS, Nashville, TN (C.C.H.); and Stanford Cardiovascular Institute, Department of Medicine, Division of Cardiology and Department of Radiology, Stanford University School of Medicine, CA (J.C.W.).
authors.forename: Charles H
authors.initials: CH
authors.lastname: Williams
authors.affiliation: From the Departments of Pharmacology (T.K.F.), Cardiovascular Medicine (Y.W.C., J.E.H., C.C.L., C.C.H.), and Department of Medicine, Divisions of Cardiovascular Medicine and Clinical Pharmacology, Oates Institute for Experimental Therapeutics (L.W., B.C.K.), Department of Molecular Physiology and Biophysics (A.G.C.), Vanderbilt University School of Medicine, Nashville, TN; Departments of Cell and Developmental Biology (C.H.W.) and Biomedical Engineering (N.B., W.D.M.), Vanderbilt University, Nashville, TN; Research Medicine, Veterans Affairs TVHS, Nashville, TN (C.C.H.); and Stanford Cardiovascular Institute, Department of Medicine, Division of Cardiology and Department of Radiology, Stanford University School of Medicine, CA (J.C.W.).
authors.forename: Young Wook
authors.initials: YW
authors.lastname: Chun
authors.affiliation: From the Departments of Pharmacology (T.K.F.), Cardiovascular Medicine (Y.W.C., J.E.H., C.C.L., C.C.H.), and Department of Medicine, Divisions of Cardiovascular Medicine and Clinical Pharmacology, Oates Institute for Experimental Therapeutics (L.W., B.C.K.), Department of Molecular Physiology and Biophysics (A.G.C.), Vanderbilt University School of Medicine, Nashville, TN; Departments of Cell and Developmental Biology (C.H.W.) and Biomedical Engineering (N.B., W.D.M.), Vanderbilt University, Nashville, TN; Research Medicine, Veterans Affairs TVHS, Nashville, TN (C.C.H.); and Stanford Cardiovascular Institute, Department of Medicine, Division of Cardiology and Department of Radiology, Stanford University School of Medicine, CA (J.C.W.).
authors.forename: Jonathan E
authors.initials: JE
authors.lastname: Hempel
authors.affiliation: From the Departments of Pharmacology (T.K.F.), Cardiovascular Medicine (Y.W.C., J.E.H., C.C.L., C.C.H.), and Department of Medicine, Divisions of Cardiovascular Medicine and Clinical Pharmacology, Oates Institute for Experimental Therapeutics (L.W., B.C.K.), Department of Molecular Physiology and Biophysics (A.G.C.), Vanderbilt University School of Medicine, Nashville, TN; Departments of Cell and Developmental Biology (C.H.W.) and Biomedical Engineering (N.B., W.D.M.), Vanderbilt University, Nashville, TN; Research Medicine, Veterans Affairs TVHS, Nashville, TN (C.C.H.); and Stanford Cardiovascular Institute, Department of Medicine, Division of Cardiology and Department of Radiology, Stanford University School of Medicine, CA (J.C.W.).
authors.forename: Nathaniel
authors.initials: N
authors.lastname: Bloodworth
authors.affiliation: From the Departments of Pharmacology (T.K.F.), Cardiovascular Medicine (Y.W.C., J.E.H., C.C.L., C.C.H.), and Department of Medicine, Divisions of Cardiovascular Medicine and Clinical Pharmacology, Oates Institute for Experimental Therapeutics (L.W., B.C.K.), Department of Molecular Physiology and Biophysics (A.G.C.), Vanderbilt University School of Medicine, Nashville, TN; Departments of Cell and Developmental Biology (C.H.W.) and Biomedical Engineering (N.B., W.D.M.), Vanderbilt University, Nashville, TN; Research Medicine, Veterans Affairs TVHS, Nashville, TN (C.C.H.); and Stanford Cardiovascular Institute, Department of Medicine, Division of Cardiology and Department of Radiology, Stanford University School of Medicine, CA (J.C.W.).
authors.forename: W David
authors.initials: WD
authors.lastname: Merryman
authors.affiliation: From the Departments of Pharmacology (T.K.F.), Cardiovascular Medicine (Y.W.C., J.E.H., C.C.L., C.C.H.), and Department of Medicine, Divisions of Cardiovascular Medicine and Clinical Pharmacology, Oates Institute for Experimental Therapeutics (L.W., B.C.K.), Department of Molecular Physiology and Biophysics (A.G.C.), Vanderbilt University School of Medicine, Nashville, TN; Departments of Cell and Developmental Biology (C.H.W.) and Biomedical Engineering (N.B., W.D.M.), Vanderbilt University, Nashville, TN; Research Medicine, Veterans Affairs TVHS, Nashville, TN (C.C.H.); and Stanford Cardiovascular Institute, Department of Medicine, Division of Cardiology and Department of Radiology, Stanford University School of Medicine, CA (J.C.W.).
authors.forename: Chee Chew
authors.initials: CC
authors.lastname: Lim
authors.affiliation: From the Departments of Pharmacology (T.K.F.), Cardiovascular Medicine (Y.W.C., J.E.H., C.C.L., C.C.H.), and Department of Medicine, Divisions of Cardiovascular Medicine and Clinical Pharmacology, Oates Institute for Experimental Therapeutics (L.W., B.C.K.), Department of Molecular Physiology and Biophysics (A.G.C.), Vanderbilt University School of Medicine, Nashville, TN; Departments of Cell and Developmental Biology (C.H.W.) and Biomedical Engineering (N.B., W.D.M.), Vanderbilt University, Nashville, TN; Research Medicine, Veterans Affairs TVHS, Nashville, TN (C.C.H.); and Stanford Cardiovascular Institute, Department of Medicine, Division of Cardiology and Department of Radiology, Stanford University School of Medicine, CA (J.C.W.).
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Wu
authors.affiliation: From the Departments of Pharmacology (T.K.F.), Cardiovascular Medicine (Y.W.C., J.E.H., C.C.L., C.C.H.), and Department of Medicine, Divisions of Cardiovascular Medicine and Clinical Pharmacology, Oates Institute for Experimental Therapeutics (L.W., B.C.K.), Department of Molecular Physiology and Biophysics (A.G.C.), Vanderbilt University School of Medicine, Nashville, TN; Departments of Cell and Developmental Biology (C.H.W.) and Biomedical Engineering (N.B., W.D.M.), Vanderbilt University, Nashville, TN; Research Medicine, Veterans Affairs TVHS, Nashville, TN (C.C.H.); and Stanford Cardiovascular Institute, Department of Medicine, Division of Cardiology and Department of Radiology, Stanford University School of Medicine, CA (J.C.W.). charles.c.hong@vanderbilt.edu bjorn.knollmann@vanderbilt.edu.
authors.forename: Björn C
authors.initials: BC
authors.lastname: Knollmann
authors.affiliation: From the Departments of Pharmacology (T.K.F.), Cardiovascular Medicine (Y.W.C., J.E.H., C.C.L., C.C.H.), and Department of Medicine, Divisions of Cardiovascular Medicine and Clinical Pharmacology, Oates Institute for Experimental Therapeutics (L.W., B.C.K.), Department of Molecular Physiology and Biophysics (A.G.C.), Vanderbilt University School of Medicine, Nashville, TN; Departments of Cell and Developmental Biology (C.H.W.) and Biomedical Engineering (N.B., W.D.M.), Vanderbilt University, Nashville, TN; Research Medicine, Veterans Affairs TVHS, Nashville, TN (C.C.H.); and Stanford Cardiovascular Institute, Department of Medicine, Division of Cardiology and Department of Radiology, Stanford University School of Medicine, CA (J.C.W.). charles.c.hong@vanderbilt.edu bjorn.knollmann@vanderbilt.edu.
authors.forename: Charles C
authors.initials: CC
authors.lastname: Hong
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D003094:Collagen / Q000008:administration & dosage; D004338:Drug Combinations; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000502:physiology*; D007797:Laminin / Q000008:administration & dosage; D009200:Myocardial Contraction / Q000187:drug effects / Q000502:physiology; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000502:physiology*; D039904:Pluripotent Stem Cells / Q000187:drug effects / Q000502:physiology; D011509:Proteoglycans / Q000008:administration & dosage
chemical_list: D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen
keywords: excitation contraction coupling; matrigel; myocytes, cardiac; pluripotent stem cells; stem cells