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DZHK (German Centre for Cardiovascular Research)

14 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Bioluminescence Imaging for Assessment of Immune Responses Following Implantation of Engineered Heart Tissue (EHT)
Lenard Conradi 1,2, Christiane Pahrmann 1, Stephanie Schmidt 1, Tobias Deuse 1,3, Arne Hansen 2, Alexandra Eder 2, Hermann Reichenspurner 1, Robert C. Robbins 3, Thomas Eschenhagen 2, Sonja Schrepfer 1,3
1Transplant and Stem Cell Immunobiology Lab (TSI) and CVRC, University Hospital Hamburg, University Heart Center Hamburg, 2Department of Experimental and Clinical Pharmacology and Toxicology, University Heart Center Hamburg, 3CT Surgery, Stanford University School of Medicine

This video demonstrates the use of in vivo bioluminescence imaging to study immune responses after implantation of Engineered Heart Tissue (EHT) in rats.

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Biology

Isolation of Human Atrial Myocytes for Simultaneous Measurements of Ca2+ Transients and Membrane Currents
Niels Voigt *1,2, Xiao-Bo Zhou *2, Dobromir Dobrev 1,2
1Institute of Pharmacology, University of Duisburg-Essen , 2Division of Experimental Cardiology, University of Heidelberg

We describe the isolation of human atrial myocytes which can be used for intracellular Ca2+ measurements in combination with electrophysiological patch-clamp studies.

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JoVE Core

Automated Contraction Analysis of Human Engineered Heart Tissue for Cardiac Drug Safety Screening
Ingra Mannhardt 1, Umber Saleem 1, Anika Benzin 1, Thomas Schulze 1, Birgit Klampe 1, Thomas Eschenhagen 1, Arne Hansen 1
1Department of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf and DZHK (German Center for Cardiovascular Research)

Here, we show the generation of human engineered heart tissue from induced pluripotent stem cells (hiPSC)-derived cardiomyocytes. We present a method to analyze contraction force and exemplary alteration of contraction pattern by the hERG channel inhibitor E-4031. This method shows high level of robustness and suitability for cardiac drug screening.

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Medicine

Impact of Intracardiac Neurons on Cardiac Electrophysiology and Arrhythmogenesis in an Ex Vivo Langendorff System
Christiane Jungen 1,2, Katharina Scherschel 1,2, Nadja I. Bork 2,3, Pawel Kuklik 1, Christian Eickholt 1, Helge Kniep 1, Niklas Klatt 1,2, Stephan Willems 1,2, Viacheslav O. Nikolaev 2,3, Christian Meyer 1,2
1Department of Cardiology-Electrophysiology, cNEP (cardiac Neuro- and Electrophysiology research group), University Heart Center, University Hospital Hamburg-Eppendorf, 2DZHK (German Center for Cardiovascular Research), 3Institute of Experimental Cardiovascular Research, University Medical Center Hamburg-Eppendorf

Here, we present a protocol for the modulation of the intracardiac autonomic nervous system and the assessment of its influence on basic electrophysiology, arrhythmogenesis, and cAMP dynamics using an ex vivo Langendorff setup.

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Medicine

Implantation of hiPSC-derived Cardiac-muscle Patches after Myocardial Injury in a Guinea Pig Model
Liesa Castro 1,2, Birgit Geertz 3, Marina Reinsch 2,3, Bülent Aksehirlioglu 3, Arne Hansen 2,3, Thomas Eschenhagen 2,3, Hermann Reichenspurner 1,2, Florian Weinberger *2,3, Simon Pecha *1,2
1Department of Cardiovascular Surgery, University Heart Center Hamburg, 2partner site Hamburg/Kiel/Lübeck, German Centre for Cardiovascular Research (DZHK), 3Department of Experimental Pharmacology and Toxicology, Cardiovascular ResearchCenter, University Medical Center Hamburg-Eppendorf

Here we present a protocol for the induction of left ventricular cryoinjury followed by the implantation of a cardiac muscle patch, derived from human iPS-cell cardiomyocytes in a guinea pig model.

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Behavior

Visualization of Intensity Levels to Reduce the Gap Between Self-Reported and Directly Measured Physical Activity
Lisa Voigt 1,4, Antje Ullrich 1,4, Ulrike Siewert-Markus 1,2,4, Marcus Dörr 3,4, Ulrich John 1,4, Sabina Ulbricht 1,4
1Institute of Social Medicine and Prevention, University Medicine Greifswald, 2Institute for Medical Psychology, University Medicine Greifswald, 3Department of Internal Medicine B, University Medicine Greifswald, 4partner site Greifswald, DZHK (German Centre for Cardiovascular Research)

This protocol describes a randomized controlled trial as a method to test the effect of a video demonstration on the intra-individual difference between self-reported and accelerometer-based moderate-to-vigorous physical activity.

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Immunology and Infection

An Advanced Murine Model for Nonalcoholic Steatohepatitis in Association with Type 2 Diabetes
Julia Sbierski-Kind 1,2,3,4, Katharina Schmidt-Bleek 3,6, Mathias Streitz 3,5, Jonas Kath 3,5, Joachim Spranger *1,2,4, Hans-Dieter Volk *2,3,5
1Department of Endocrinology & Metabolism Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, 2Berlin Institute of Health (BIH), 3Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, 4DZHK (German Centre for Cardiovascular Research), 5Institute of Medical Immunology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, 6Julius Wolff Institute (JWI) and Center for Musculoskeletal Surgery, Charité - Universitätsmedizin Berlin, Berlin, Germany, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin

A simple and reliable diet-induced rodent animal model for nonalcoholic steatohepatitis (NASH) is described, achieved through non-SPF housing of the animals and administration of a specific high-fat diet. We describe identification of hepatic and adipose immune cell subsets to recapitulate human immunological conditions by exposing mice to environmental germs.

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Developmental Biology

Magnetic Adjustment of Afterload in Engineered Heart Tissues
Benjamin Becker *1,2, Marita L. Rodriguez *1,2, Tessa R. Werner 1,2, Justus Stenzig 1,2, Thomas Eschenhagen 1,2, Marc N. Hirt 1,2
1Institute of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, 2DZHK (German Centre for Cardiovascular Research)

This protocol provides detailed methods describing the fabrication and implementation of a magnetics-based afterload tuning platform for engineered heart tissues.

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Bioengineering

Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes
Fitzwilliam Seibertz 1,2, Martyn Reynolds 3, Niels Voigt 1,2,4
1Institute of Pharmacology and Toxicology, University Medical Center Goettingen, Goettingen, Germany, 2DZHK (German Center for Cardiovascular Research), Partner Site Goettingen, Germany, 3Cairn Research Ltd, Faversham, United Kingdom, 4Cluster of Excellence "Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells" (MBExC), University of Goettingen, Germany

Here we describe optical acquisition and characterization of action potentials from induced pluripotent stem cell derived cardiomyocytes using a high-speed modular photometry system.

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Biology

Isolation of High Quality Murine Atrial and Ventricular Myocytes for Simultaneous Measurements of Ca2+ Transients and L-Type Calcium Current
Philipp Tomsits *1,2,3, Dominik Schüttler *1,2,3, Stefan Kääb 1,2, Sebastian Clauss *1,2,3, Niels Voigt *4,5,6
1Department of Medicine I, University Hospital Munich, Campus Großhadern, Ludwig-Maximilians University Munich (LMU), 2Partner Site Munich, Munich Heart Alliance (MHA), DZHK (German Centre for Cardiovascular Research), 3Walter Brendel Center of Experimental Medicine, Ludwig-Maximilians University Munich (LMU), 4Institute of Pharmacology and Toxicology, University Medical Center Göttingen, 5Partner Site Göttingen, DZHK (German Centre for Cardiovascular Research), 6Cluster of Excellence "Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells" (MBExC), University of Göttingen

Mouse models allow studying key mechanisms of arrhythmogenesis. For this purpose, high quality cardiomyocytes are necessary to perform patch-clamp measurements. Here, a method to isolate murine atrial and ventricular myocytes via retrograde enzyme-based Langendorff perfusion, which allows simultaneous measurements of calcium-transients and L-type calcium current, is described.

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Medicine

Location, Dissection, and Analysis of the Murine Stellate Ganglion
Katharina Scherschel 1,2,3, Hanna Bräuninger 2,4, Klara Glufke 2, Christiane Jungen 2,4,5, Nikolaj Klöcker 3, Christian Meyer 1,2,3
1Division of Cardiology, EVK Düsseldorf, cNEP, cardiac Neuro- and Electrophysiology Research Consortium, 2DZHK (German Centre for Cardiovascular Research), 3Institute of Neural and Sensory Physiology, Medical Faculty, Heinrich Heine University Düsseldorf, 4Clinic for Cardiology, University Heart & Vascular Centre, University Hospital Hamburg-Eppendorf, 5Department of Cardiology, Leiden University Medical Center

Pathophysiological changes in the cardiac autonomic nervous system, especially in its sympathetic branch, contribute to the onset and maintenance of ventricular arrhythmias. In the present protocol, we show how to characterize murine stellate ganglia to improve the understanding of the underlying molecular and cellular processes.

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Medicine

Isolation and Culture of Resident Cardiac Macrophages from the Murine Sinoatrial and Atrioventricular Node
Ruibing Xia 1,2,3, Simone Loy 1, Stefan Kääb 1,3, Anna Titova 1, Christian Schulz 1,2,3, Steffen Massberg 1,2,3, Sebastian Clauss 1,2,3
1University Hospital Munich, Department of Medicine I, Ludwig-Maximilian-Unversity Munich (LMU), 2Insitute of Surgical Research at the Walter Brendel Center of Experimental Medicine, University Hospital Munich, Ludwig-Maximilians-University (LMU), 3German Center for Cardiovascular Research (DZHK), Partner Site Munich, Munich Heart Alliance (MHA)

The protocol presented here provides a step-by-step approach for the isolation of cardiac resident macrophages from the sinoatrial node (SAN) and atrioventricular node (AVN) region of mouse hearts.

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Medicine

Analyzing Long-Term Electrocardiography Recordings to Detect Arrhythmias in Mice
Philipp Tomsits 1,2,3, Kavi Raj Chataut 1,2, Aparna Sharma Chivukula 1,2, Li Mo 1,2, Ruibing Xia 1,2, Dominik Schüttler 1,2,3, Sebastian Clauss 1,2,3
1Department of Medicine I, University Hospital Munich, Campus Großhadern, Ludwig-Maximilians University Munich (LMU), 2DZHK (German Centre for Cardiovascular Research), Partner Site Munich, Munich Heart Alliance (MHA), 3Walter Brendel Centre of Experimental Medicine, Ludwig-Maximilians University Munich (LMU)

Here we present a step-by-step protocol for a semiautomated approach to analyze murine long-term electrocardiography (ECG) data for basic ECG parameters and common arrhythmias. Data are obtained by implantable telemetry transmitters in living and awake mice and analyzed using Ponemah and its analysis modules.

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Medicine

Real-Time Electrocardiogram Monitoring During Treadmill Training in Mice
Philipp Tomsits *1,2,4, Aparna Sharma Chivukula *1,2, Kavi Raj Chataut *1,2, Agus Simahendra 3, Ludwig T. Weckbach 1,2,3, Stefan Brunner 1, Sebastian Clauss 1,2,4
1Department of Medicine I, University Hospital Munich, Ludwig-Maximilians University Munich (LMU), 2DZHK (German Centre for Cardiovascular Research), Partner Site Munich, Munich Heart Alliance (MHA), 3Institute of Cardiovascular Physiology and Pathophysiology, Biomedical Center, Ludwig-Maximilians-University, 4Institute of Surgical Research at the Walter-Brendel-Centre of Experimental Medicine, University Hospital Munich, Ludwig-Maximilians University Munich (LMU)

Electrocardiogram (ECG) is the key variable to understanding cardiac electrophysiology. Physical exercise has beneficial effects but may also be harmful in the context of cardiovascular diseases. This manuscript provides a method of recording real-time ECG during exercise, which can serve to investigate its effects on cardiac electrophysiology in mice.

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