title: Isolation and Mechanical Measurements of Myofibrils from Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.
abstract: Tension production and contractile properties are poorly characterized aspects of excitation-contraction coupling of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Previous approaches have been limited due to the small size and structural immaturity of early-stage hiPSC-CMs. We developed a substrate nanopatterning approach to produce hiPSC-CMs in culture with adult-like dimensions, T-tubule-like structures, and aligned myofibrils. We then isolated myofibrils from hiPSC-CMs and measured the tension and kinetics of activation and relaxation using a custom-built apparatus with fast solution switching. The contractile properties and ultrastructure of myofibrils more closely resembled human fetal myofibrils of similar gestational age than adult preparations. We also demonstrated the ability to study the development of contractile dysfunction of myofibrils from a patient-derived hiPSC-CM cell line carrying the familial cardiomyopathy MYH7 mutation (E848G). These methods can bring new insights to understanding cardiomyocyte maturation and developmental mechanical dysfunction of hiPSC-CMs with cardiomyopathic mutations.
authors.affiliation: Division of Physiology, Department of Experimental and Clinical Medicine, University of Florence, Viale Morgagni 63, 50134 Florence, Italy. Electronic address: jose.pioner@unisi.it.
authors.forename: Josè Manuel
authors.initials: JM
authors.lastname: Pioner
authors.affiliation: Bioengineering, University of Washington, Seattle, WA 98109, USA.
authors.forename: Alice W
authors.initials: AW
authors.lastname: Racca
authors.affiliation: Bioengineering, University of Washington, Seattle, WA 98109, USA.
authors.forename: Jordan M
authors.initials: JM
authors.lastname: Klaiman
authors.affiliation: Medicine, Division of Cardiology, University of Washington, Seattle, WA 98195, USA.
authors.forename: Kai-Chun
authors.initials: KC
authors.lastname: Yang
authors.affiliation: Rehabilitation Medicine, University of Washington, Seattle, WA 98109, USA.
authors.forename: Xuan
authors.initials: X
authors.lastname: Guan
authors.affiliation: Pathology, University of Washington, Seattle, WA 98109, USA.
authors.forename: Lil
authors.initials: L
authors.lastname: Pabon
authors.affiliation: Pathology, University of Washington, Seattle, WA 98109, USA.
authors.forename: Veronica
authors.initials: V
authors.lastname: Muskheli
authors.affiliation: Bioengineering, University of Washington, Seattle, WA 98109, USA.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Zaunbrecher
authors.affiliation: Bioengineering, University of Washington, Seattle, WA 98109, USA.
authors.forename: Jesse
authors.initials: J
authors.lastname: Macadangdang
authors.affiliation: Medicine, Division of Cardiology, University of Colorado, Denver, CO 80262, USA.
authors.forename: Mark Y
authors.initials: MY
authors.lastname: Jeong
authors.affiliation: Rehabilitation Medicine, University of Washington, Seattle, WA 98109, USA; Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA.
authors.forename: David L
authors.initials: DL
authors.lastname: Mack
authors.affiliation: Rehabilitation Medicine, University of Washington, Seattle, WA 98109, USA; Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA.
authors.forename: Martin K
authors.initials: MK
authors.lastname: Childers
authors.affiliation: Bioengineering, University of Washington, Seattle, WA 98109, USA.
authors.forename: Deok-Ho
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Division of Physiology, Department of Experimental and Clinical Medicine, University of Florence, Viale Morgagni 63, 50134 Florence, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Tesi
authors.affiliation: Division of Physiology, Department of Experimental and Clinical Medicine, University of Florence, Viale Morgagni 63, 50134 Florence, Italy.
authors.forename: Corrado
authors.initials: C
authors.lastname: Poggesi
authors.affiliation: Bioengineering, University of Washington, Seattle, WA 98109, USA; Medicine, Division of Cardiology, University of Washington, Seattle, WA 98195, USA; Pathology, University of Washington, Seattle, WA 98109, USA; Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA; Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA.
authors.forename: Charles E
authors.initials: CE
authors.lastname: Murry
authors.affiliation: Bioengineering, University of Washington, Seattle, WA 98109, USA; Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA; Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Regnier
mesh_terms: D001696:Biomechanical Phenomena; D024722:Cardiac Myosins / Q000235:genetics; D009202:Cardiomyopathies / Q000235:genetics / Q000503:physiopathology; D002454:Cell Differentiation; D002460:Cell Line; D056966:Excitation Contraction Coupling / Q000502:physiology*; D015870:Gene Expression; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology*; D007700:Kinetics; D009154:Mutation; D032383:Myocytes, Cardiac / Q000166:cytology / Q000502:physiology*; D009210:Myofibrils / Q000502:physiology* / Q000648:ultrastructure; D018995:Myosin Heavy Chains / Q000235:genetics; D049329:Nanostructures / Q000737:chemistry; D061251:Primary Cell Culture
chemical_list: C501641:MYH7 protein, human; D024722:Cardiac Myosins; D018995:Myosin Heavy Chains