Isolation and Mechanical Measurements of Myofibrils from Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes
| Autor: | Kai Chun Yang, Rebecca J. Zaunbrecher, Chiara Tesi, Mark Y. Jeong, Deok Ho Kim, Josè Manuel Pioner, Jordan M. Klaiman, Alice Ward Racca, David L. Mack, Charles E. Murry, Martin K. Childers, Lil Pabon, Corrado Poggesi, Michael Regnier, Veronica Muskheli, Xuan Guan, Jesse Macadangdang |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Cellular differentiation Induced Pluripotent Stem Cells Primary Cell Culture Gene Expression Biology Biochemistry Sarcomere Article Cell Line 03 medical and health sciences Myofibrils Myosin Genetics Humans Myocyte Myocytes Cardiac Induced pluripotent stem cell lcsh:QH301-705.5 Excitation Contraction Coupling health care economics and organizations lcsh:R5-920 Myosin Heavy Chains Cell Differentiation Cell Biology Anatomy Biomechanical Phenomena Nanostructures 3. Good health Cell biology Kinetics 030104 developmental biology lcsh:Biology (General) Cell culture STEM CELLS CARDIOMYOCYTES SARCOMERE CARDIOMYOPATHIES Mutation Stem cell Cardiomyopathies Myofibril lcsh:Medicine (General) Cardiac Myosins Developmental Biology |
| Zdroj: | Stem Cell Reports, Vol 6, Iss 6, Pp 885-896 (2016) Stem Cell Reports |
| Popis: | Summary 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. Highlights • The contractile properties of hiPSC-CM myofibrils have not been previously studied • hiPSC-CMs cultured on nanopatterned surfaces develop elongated, aligned myofibrils • hiPSC-CMs myofibrils have contractile properties similar to human fetal myofibrils • hiPSC-CMs can be used to study development of genetically based cardiac diseases In this article, Pioner and colleagues reported contractile properties of isolated myofibrils from hiPSC-CMs with highly mature morphology. This approach permits quantitative assessment of maturation and contractile properties of hiPSC-CMs and can be used to study the development of contractile dysfunction in genetically based cardiac diseases. The authors present a patient-derived cell line carrying a novel familial cardiomyopathy MYH7 mutation (E848G). |
| Databáze: | OpenAIRE |
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