In Vitro Organization of Dissociated Rat Cardiac Cells Into Beating Three-Dimensional Structures

Halbert, Seymour P.; Bruderer, Ruth; Lin, Tsue-Ming

doi:10.1084/jem.133.4.677

Cited by 31 publications

(13 citation statements)

References 19 publications

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“…Scaffold-based cardiac constructs promote the linear alignment of cardiomyocytes and, by direct attachment to sensors, have been used to measure contractile force and other physiologic parameters (24). The hanging drop method for cell culture, as we have used here, is actually an older concept and even the use of cardiomyocytes to produce "mini hearts" dates back to the early seventies (25). A couple of pioneering studies of the last decade have provided technological innovations and a better understanding of this type of 3D culture and its potential for applications in tissue engineering and drug development (5,26,27).…”

Section: Discussionmentioning

confidence: 99%

Development and Characterization of a Scaffold-Free 3D Spheroid Model of Induced Pluripotent Stem Cell-Derived Human Cardiomyocytes

Beauchamp

Moritz

Kelm

et al. 2015

Tissue Engineering Part C: Methods

152

121

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Cardiomyocytes (CMs) are terminally differentiated cells in the adult heart, and ischemia and cardiotoxic compounds can lead to cell death and irreversible decline of cardiac function. As testing platforms, isolated organs and primary cells from rodents have been the standard in research and toxicology, but there is a need for better models that more faithfully recapitulate native human biology. Hence, a new in vitro model comprising the advantages of 3D cell culture and the availability of induced pluripotent stem cells (iPSCs) of human origin was developed and characterized. Human CMs derived from iPSCs were studied in standard 2D culture and as cardiac microtissues (MTs) formed in hanging drops. Two-dimensional cultures were examined using immunofluorescence microscopy and western blotting, while the cardiac MTs were subjected to immunofluorescence, contractility, and pharmacological investigations. iPSC-derived CMs in 2D culture showed well-formed myofibrils, cell-cell contacts positive for connexin-43, and other typical cardiac proteins. The cells reacted to prohypertrophic growth factors with a substantial increase in myofibrils and sarcomeric proteins. In hanging drop cultures, iPSC-derived CMs formed spheroidal MTs within 4 days, showing a homogeneous tissue structure with well-developed myofibrils extending throughout the whole spheroid without a necrotic core. MTs showed spontaneous contractions for more than 4 weeks that were recorded by optical motion tracking, sensitive to temperature and responsive to electrical pacing. Contractile pharmacology was tested with several agents known to modulate cardiac rate and viability. Calcium transients underlay the contractile activity and were also responsive to electrical stimulation, caffeine-induced Ca(2+) release, and extracellular calcium levels. A three-dimensional culture using iPSC-derived human CMs provides an organoid human-based cellular platform that is free of necrosis and recapitulates vital cardiac functionality, thereby providing a new and promising relevant model for the evaluation and development of new therapies and detection of cardiotoxicity.

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Section: Discussionmentioning

confidence: 99%

Development and Characterization of a Scaffold-Free 3D Spheroid Model of Induced Pluripotent Stem Cell-Derived Human Cardiomyocytes

Beauchamp

Moritz

Kelm

et al. 2015

Tissue Engineering Part C: Methods

152

121

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“…It is common for monolayers and multilayers of cardiac muscle cells and smooth muscle cells, which contract spontaneously, to pull loose from the substrate and form three-dimensional assemblies of reaggregated cells, which usually assume a more-or-less spherical shape eventually. The formation of such spherical reaggregates can be expedited by growing the cells on a substrate to which they adhere poorly, such as cellophane (14). Spherical reaggregates can also be formed for other cell types besides muscle by spinning the enzymedissociated cells in a gyrotatory shaker (15).…”

Section: Types Of Culturesmentioning

confidence: 99%

“…The cells can be plated into glass culture dishes containing cellophane squares on the bottom. Since the cells do not adhere very well to cellophane, they pull free and form small spherical reaggregates (0.1-0.5 mm) spontaneously (14,40). Alternatively, the cells can be placed into Ehrlenmeyer flasks and rotated on a gyrotatory shaker for about 48 hr (15).…”

Section: Spherical Reaggregatesmentioning

confidence: 99%

Cultured Heart Cell Reaggregate Model for Studying Cardiac Toxicology

Sperelakis¹

1978

Environmental Health Perspectives

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JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms of scholarship. For more information about JSTOR, please contact support@jstor.org.. The National Institute of Environmental Health Sciences (NIEHS) andBrogan & Partners are collaborating with JSTOR to digitize, preserve and extend access to Environmental Health Perspectives. This review represents a summary of the technique of using cultured heart cells as a model system for studying the physiology, pharmacology, biochemistry and toxicology of myocardial cells. The general techniques and types of culture preparations commonly used are given and some of the advantages and disadvantages of working with cultured heart cells are summarized.

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“…The ability of isolated ventricular myocytes in culture to spontaneously organize into 3D structures has long been recognized (27). Moreover, a number of 3D tissue culture techniques were developed as biological systems for studying cell-cell interactions, electrical coupling, and extracellular matrix interactions (28–30).…”

Section: Discussionmentioning

confidence: 99%

Engineered Electrical Conduction Tract Restores Conduction in Complete Heart Block

Cingolani¹,

Ionta

Cheng³

et al. 2014

Journal of the American College of Cardiology

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BACKGROUND Cardiac electrical conduction delays and blocks cause rhythm disturbances such as complete heart block, which can be fatal. Standard of care relies on electronic devices to restore synchrony artificially. We sought to create a new modality to treat such disorders by engineering electrical conduction tracts designed to propagate electrical impulses. OBJECTIVES We sought to create a new approach to treat cardiac conduction disorders using engineered electrical conduction tracts (EECT). METHODS Paramagnetic beads were conjugated with an antibody to γ-sarcoglycan, a cardiomyocyte cell-surface antigen, and mixed with freshly-isolated neonatal rat ventricular cardiomyocytes (NRVMs). A magnetic field was used to pattern a linear EECT. RESULTS In an in vitro model of conduction block, EECT, patterned so as to connect 2 independently beating NRVM monolayers, achieved coordinated electrical activity, with action potentials propagating from 1 region to the other via EECT. Spiking EECT with heart-derived stromal cells yielded stable structures with highly reproducible conduction velocities. Transplantation of EECT in vivo restored atrio-ventricular conduction in a rat model of complete heart block. CONCLUSIONS A tissue-engineered electrical conduction tract can re-establish electrical conduction in the heart. This novel approach could, in principle, be used, not only to treat cardiac arrhythmias, but also to repair other organs.

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In Vitro Organization of Dissociated Rat Cardiac Cells Into Beating Three-Dimensional Structures

Cited by 31 publications

References 19 publications

Development and Characterization of a Scaffold-Free 3D Spheroid Model of Induced Pluripotent Stem Cell-Derived Human Cardiomyocytes

Development and Characterization of a Scaffold-Free 3D Spheroid Model of Induced Pluripotent Stem Cell-Derived Human Cardiomyocytes

Cultured Heart Cell Reaggregate Model for Studying Cardiac Toxicology

Engineered Electrical Conduction Tract Restores Conduction in Complete Heart Block

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