title: Extracellular Matrix-Mediated Maturation of Human Pluripotent Stem Cell-Derived Cardiac Monolayer Structure and Electrophysiological Function.
abstract: BACKGROUND
  Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) monolayers generated to date display an immature embryonic-like functional and structural phenotype that limits their utility for research and cardiac regeneration. In particular, the electrophysiological function of hPSC-CM monolayers and bioengineered constructs used to date are characterized by slow electric impulse propagation velocity and immature action potential profiles.
  
  
  RESULTS
  Here, we have identified an optimal extracellular matrix for significant electrophysiological and structural maturation of hPSC-CM monolayers. hPSC-CM plated in the optimal extracellular matrix combination have impulse propagation velocities ≈2× faster than previously reported (43.6±7.0 cm/s; n=9) and have mature cardiomyocyte action potential profiles, including hyperpolarized diastolic potential and rapid action potential upstroke velocity (146.5±17.7 V/s; n=5 monolayers). In addition, the optimal extracellular matrix promoted hypertrophic growth of cardiomyocytes and the expression of key mature sarcolemmal (SCN5A, Kir2.1, and connexin43) and myofilament markers (cardiac troponin I). The maturation process reported here relies on activation of integrin signaling pathways: neutralization of β1 integrin receptors via blocking antibodies and pharmacological blockade of focal adhesion kinase activation prevented structural maturation.
  
  
  CONCLUSIONS
  Maturation of human stem cell-derived cardiomyocyte monolayers is achieved in a 1-week period by plating cardiomyocytes on PDMS (polydimethylsiloxane) coverslips rather than on conventional 2-dimensional cell culture formats, such as glass coverslips or plastic dishes. Activation of integrin signaling and focal adhesion kinase is essential for significant maturation of human cardiac monolayers.
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.). toddherr@umich.edu.
authors.forename: Todd J
authors.initials: TJ
authors.lastname: Herron
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: Andre Monteiro Da
authors.initials: AM
authors.lastname: Rocha
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: Katherine F
authors.initials: KF
authors.lastname: Campbell
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: Daniela
authors.initials: D
authors.lastname: Ponce-Balbuena
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: B Cicero
authors.initials: BC
authors.lastname: Willis
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: Guadalupe
authors.initials: G
authors.lastname: Guerrero-Serna
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: Qinghua
authors.initials: Q
authors.lastname: Liu
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: Matt
authors.initials: M
authors.lastname: Klos
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: Hassan
authors.initials: H
authors.lastname: Musa
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: Manuel
authors.initials: M
authors.lastname: Zarzoso
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: Alexandra
authors.initials: A
authors.lastname: Bizy
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: Jamie
authors.initials: J
authors.lastname: Furness
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: Justus
authors.initials: J
authors.lastname: Anumonwo
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: Sergey
authors.initials: S
authors.lastname: Mironov
authors.affiliation: From the Center for Arrhythmia Research, Department of Internal Medicine, Cardiovascular Research Center, University of Michigan, Ann Arbor, MI (T.J.H., A.M.D.R., K.C., D.P.-B., B.C.W., G.G.-S., Q.L., J.F., J.A., S.M., J.J.); Department of Medicine, University of California San Diego, La Jolla, CA (M.K.); Davis Heart and Lung Research Institute, Ohio State University, Columbus, OH (H.M.); Shanxi Medical University, Zhejiang, China (Q.L.); and University of Valencia, Valencia, Spain (M.Z., A.B.).
authors.forename: José
authors.initials: J
authors.lastname: Jalife
mesh_terms: D000200:Action Potentials / Q000502:physiology; D002454:Cell Differentiation; D002460:Cell Line; D055724:Electrophysiological Phenomena / Q000502:physiology*; D005109:Extracellular Matrix / Q000502:physiology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D015398:Signal Transduction
keywords: cardiac electrophysiology; cell culture techniques; hardness; induced pluripotent stem cells; integrin alpha5beta1; myocytes, cardiac; troponin I