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Cardiac Tissue
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Methods and Protocols
Second Edition
METHODS IN MOLECULAR BIOLOGY

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School of Life and Medical Sciences
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For over 35 years, biological scientists have come to rely on the research protocols and
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Cardiac Tissue Engineering

Methods and Protocols

Second Edition

Edited by

Kareen L. K. Coulombe
School of Engineering, Brown University, Providence, RI, USA

Lauren D. Black III

Department of Biomedical Engineering, Tufts University, Medford, MA, USA
Editors
Kareen L. K. Coulombe Lauren D. Black III
School of Engineering Department of Biomedical Engineering
Brown University Tufts University
Providence, RI, USA Medford, MA, USA

ISSN 1064-3745 ISSN 1940-6029 (electronic)

Methods in Molecular Biology
ISBN 978-1-0716-2260-5 ISBN 978-1-0716-2261-2 (eBook)
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Preface

Due to the limited regenerative potential of the adult human heart, development of
alternative treatment options is needed for the variety of conditions that result in the loss,
or loss of function, of contractile tissue. Perhaps the prime example of this is myocardial
infarction (MI), which results in death of tens of millions of cardiomyocytes. While this loss
is significant, many patients receive minimal intervention following MI, and their cardiac
health is managed with drugs and changes to diet and exercise with satisfactory results.
However, within 5 years, more than half of MI patients will develop heart failure, for which
the only successful late-stage treatment is heart transplantation.
In the first edition of this book, we published a wide array of protocols important for
cardiac tissue engineering research. Many of these were geared towards in vitro advancement
ahead of future clinical work. In the time since the first edition, the field has continued to
advance in two key directions. The first is a deeper understanding of human pluripotent stem
cell-derived cardiomyocytes in three-dimensional engineered microenvironments and their
use in downstream applications. The second is continued movement towards clinical thera-
pies using tissue engineering approaches in the heart in large animal models and human
clinical trials.
In this book, the second edition of Cardiac Tissue Engineering: Methods and Protocols,
an updated collection of state-of-the-art protocols in cardiac tissue engineering is presented.
These protocols demonstrate advancements in cell sourcing, assembly, and use of engi-
neered cardiac tissues, imaging and diagnostics, and applications. Platforms developed for
broad use to study development and disease in vitro enable genotype to phenotype evalua-
tion, and many are customized for contractility, arrhythmia, or heart repair in vivo using
small and large animal models. New animal models, biomaterials, and quantitative analyses
are described for broad adoption.
The diversity of research in cardiovascular development and disease has inspired the
development of a number of techniques and platforms that can be used to address an array of
questions, often leveraging the use of human pluripotent stem cell-derived cardiomyocytes
(hiPSC-CMs). Gene editing for high-throughput genetic screening using a custom CRISPR
library in cultured cardiomyocytes is described by DeLuca and Bursac. Challenges of protein
and mRNA quantification from small samples of hiPSC-CMs are described and overcome by
customized methods presented by Entcheva and colleagues to enable high-throughput
analyses.
Tissue engineering methods are diverse, enabling selection of an approach based on the
study question and design, including considerations for size, throughput, geometry, and
endpoint metrics. Methods for forming self-assembled 3D spheroids of hiPSC-CMs and
heterotypic myocardial cells are described by Matthys and McDevitt. Fabrication of myo-
cardial scaffolds and slices, integrated with electrical and mechanical stimulation, is pre-
sented by Liao and colleagues. For more complex geometries, bioprinting of a 3D ventricle
and tips for custom shapes using the freeform reversible embedding of suspended hydrogels
(FRESH) method is shared by Feinberg and colleagues. For hydrogel-based elongated

v
vi Preface

engineered heart tissues designed for contractile, structural, and transcriptional studies, the
Sniadecki group provides templates and insights for lab-made racks of silicone posts in a
standard 24-well dish format.
Fit-for-purpose platforms require alignment of the system’s biology with quantitative
readouts and has become ever-present as the field of cardiac tissue engineering plunges into
drug testing and disease modeling in vitro. Use of microelectrode arrays for high-
throughput field potential measurements in 2D plated hiPSC-CMs to assess drug responses
is detailed by Wu and colleagues. Micro-heart muscle array technology from the Huebsch
Lab enables moderate throughput in pharmacology and pharmacogenomic studies by visual
assessment of action potential (AP), calcium transient, and contractility with compatibility
for protein and gene analyses. Studies concerned with propagation velocity, or conduction
velocity, as related to arrhythmia will benefit from high-speed visual fluorescence imaging of
calcium waves and analysis algorithms presented by McCain and colleagues in an aligned 2D
cardiac platform. A heterotypic hiPSC-CM and human cardiac fibroblast self-assembled 3D
microtissue platform for arrhythmia assessment by quantitative evaluation of all phases of the
action potential is presented by Kofron, Choi, and Coulombe. While the focus of much of
the cardiac tissue engineering space is on ventricular tissue, an atrial cardiac 3D-engineered
tissue model is described by Eschenhagen, Stenzig, et al. using elastomeric posts for
auxotonic contractions with applications in atrial-specific studies of drug responses and
disease modeling. A cardiac fibrosis model based on the Biowire II platform for contractility
assessment from the Radisic group enables local high-fibroblast content to create scar-like
tissue adjacent to normal cardiac tissue for studying fibrosis and therapeutics.
As the field of cardiac tissue engineering advances towards clinical heart regeneration,
multiple approaches to remuscularize injured hearts with hiPSC/hESC-CMs are moving
towards phase I trials. Transepicardial hiPSC-CM delivery in a swine model of acute
myocardial infarction is described by Laflamme and colleagues, while defined cellular and
culture conditions in collagen-based engineered heart tissue (EHT) is provided by Zim-
mermann and colleagues for disease modeling, drug screening, and heart repair. Methods
for constructing tubular cardiac tissue from multilayered cell sheets are provided by Okano,
Sekine et al. for applications in heart failure. Finally, in a critically important analysis of
engraftment, Brandt and Mahmoud detail their methods for quantifying cardiomyocyte
proliferation and nucleation in repaired hearts via robust histological methods.
Novel therapeutics that enable in situ repair and alternative approaches for regeneration
highlight the innovation in the field of cardiac tissue engineering. Using a biomaterials
intervention, Christman and colleagues describe injectable extracellular matrix (ECM)
scaffolds that have been in use in small and large animal models for cardiac repair and initial
safety assessment in a Phase I clinical trial. Use of ECM for encapsulation of cells for
echocardiography-directed injection in rodent models is described by Shakya, Brown, and
Davis. The impact of a biomaterials-based repair strategy on the monocyte population is
described by Suuronen and colleagues using flow cytometry analyses to quantify the levels of
major leukocyte subtypes isolated from mouse hearts. Finally, a novel model of patch-based
repair is provided by Black and colleagues, where cardiovascular patches are implanted to
widen the right ventricular outflow tract in young, rapidly growing porcine hearts to
emulate congenital heart defect reconstructive surgery.
Bringing new technologies and therapies to the clinic is a challenging task, but one that
is attainable, particularly if we as a field work in collaboration. This second edition of
 Preface vii

Cardiac Tissue Engineering: Methods and Protocols aims to be your primary resource for
implementing these cutting-edge approaches in your research. With this book, we hope to
inspire advancement of cardiotoxicity assessment, drug discovery, and heart repair and
regeneration to accelerate heart health around the globe.

Providence, RI, USA Kareen L. K. Coulombe

Medford, MA, USA Lauren D. Black III
Contents

Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi

1 CRISPR Library Screening in Cultured Cardiomyocytes . . . . . . . . . . . . . . . . . . . . . 1

Sophia DeLuca and Nenad Bursac
2 Protein and mRNA Quantification in Small Samples of Human-Induced
Pluripotent Stem Cell-Derived Cardiomyocytes
in 96-Well Microplates. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Weizhen Li, Julie L. Han, and Emilia Entcheva
3 Self-Assembled Heterotypic Cardiac Spheroids from Human
Pluripotent Stem Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Oriane B. Matthys and Todd C. McDevitt
4 Acellular Myocardial Scaffolds and Slices Fabrication, and Method
for Applying Mechanical and Electrical Simulation
to Tissue Construct . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
Bo Wang, Mickey Shah, Lakiesha N. Williams, Amy L. de Jongh Curry,
Yi Hong, Ge Zhang, and Jun Liao
5 FRESH 3D Bioprinting a Ventricle-like Cardiac Construct Using
Human Stem Cell-Derived Cardiomyocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
Brian D. Coffin, Andrew R. Hudson, Andrew Lee, and Adam W. Feinberg
6 Engineered Heart Tissues for Contractile, Structural, and Transcriptional
Assessment of Human Pluripotent Stem Cell-Derived Cardiomyocytes
in a Three-Dimensional, Auxotonic Environment . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
Samantha Bremner, Alex J. Goldstein, Ty Higashi, and Nathan J. Sniadecki
7 High-Throughput Analysis of Drug Safety Responses in Induced
Pluripotent Stem Cell-Derived Cardiomyocytes
Using Multielectrode Array . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99
Nadjet Belbachir, Nathan Cunningham, and Joseph C. Wu
8 iPSC-Derived Micro-Heart Muscle for Medium-Throughput Pharmacology
and Pharmacogenomic Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
Daniel W. Simmons and Nathaniel Huebsch
9 Quantifying Propagation Velocity from Engineered Cardiac Tissues with
High-Speed Fluorescence Microscopy and Automated
Analysis Software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
Andrew P. Petersen and Megan L. McCain
10 Arrhythmia Assessment in Heterotypic Human Cardiac
Myocyte–Fibroblast Microtissues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147
Celinda M. Kofron, Bum-Rak Choi, and Kareen L. K. Coulombe
11 Human-Engineered Atrial Tissue for Studying Atrial Fibrillation . . . . . . . . . . . . . 159
Julia Krause, Marta Lemme, Ingra Mannhardt, Alexandra Eder,
B€a rbel Ulmer, Thomas Eschenhagen, and Justus Stenzig

ix
x Contents

12 Design and Fabrication of Biological Wires for Cardiac Fibrosis

Disease Modeling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175
Erika Yan Wang, Jacob Smith, and Milica Radisic
13 Methods for Transepicardial Cell Transplantation in a Swine
Myocardial Infarction Model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191
Fanny Wulkan, Rocco Romagnuolo, Beiping Qiang,
and Michael A. Laflamme
14 Defined Engineered Human Myocardium for Disease Modeling,
Drug Screening, and Heart Repair . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213
Malte Tiburcy, Tim Meyer, Pierre-Luc Satin,
and Wolfram-Hubertus Zimmermann
15 Tubular Cardiac Tissue Bioengineered from Multi-Layered Cell Sheets
for Use in the Treatment of Heart Failure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
Hidekazu Sekine and Teruo Okano
16 Quantifying Cardiomyocyte Proliferation and Nucleation to Assess
Mammalian Cardiac Regeneration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 243
Emma B. Brandt and Ahmed I. Mahmoud
17 Injectable ECM Scaffolds for Cardiac Repair . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 255
Jervaughn D. Hunter, Todd D. Johnson, Rebecca L. Braden,
and Karen L. Christman
18 Encapsulation of Pediatric Cardiac-Derived C-Kit+ Cells in Cardiac
Extracellular Matrix Hydrogel for Echocardiography-Directed
Intramyocardial Injection in Rodents. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 269
Preety Shakya, Milton E. Brown, and Michael E. Davis
19 Characterization of the Monocyte Response to Biomaterial Therapy
for Cardiac Repair . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 279
Sarah McLaughlin, David Smyth, Emilio I. Alarcon,
and Erik J. Suuronen
20 Right Ventricular Outflow Tract Surgical Resection in Young,
Large Animal Model for the Study of Alternative
Cardiovascular Patches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 299
Elizabeth C. Porter, Whitney L. Stoppel, Raymond K. Kudej,
and Lauren D. Black III

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 311
Contributors

EMILIO I. ALARCON • BioEngineering and Therapeutic Solutions (BEaTS), Division of

Cardiac Surgery, University of Ottawa Heart Institute, Ottawa, ON, Canada;
Department of Biochemistry, Microbiology, and Immunology, University of Ottawa,
Ottawa, ON, Canada
NADJET BELBACHIR • Stanford Cardiovascular Institute, Stanford University, Stanford, CA,
USA; Division of Cardiovascular Medicine, Department of Medicine, Stanford University,
Stanford, CA, USA
LAUREN D. BLACK III • Cellular, Molecular and Developmental Biology Program,
Graduate School of Biomedical Sciences, Tufts University, Boston, MA, USA; Department
of Biomedical Engineering, Tufts University, Medford, MA, USA
REBECCA L. BRADEN • Sanford Consortium for Regenerative Medicine, Department of
Bioengineering, University of California San Diego, La Jolla, CA, USA
EMMA B. BRANDT • Department of Cell and Regenerative Biology, University of Wisconsin-
Madison School of Medicine and Public Health, Madison, WI, USA
SAMANTHA BREMNER • Institute for Stem Cell and Regenerative Medicine, University of
Washington, Seattle, WA, USA; Department of Bioengineering, University of Washington,
Seattle, WA, USA
MILTON E. BROWN • Wallace H. Coulter Department of Biomedical Engineering, Georgia
Institute of Technology and Emory University School of Medicine, Atlanta, GA, USA;
Division of Cardiology, Department of Medicine, Emory University School of Medicine,
Atlanta, GA, USA
NENAD BURSAC • Department of Biomedical Engineering, Duke University, Durham, NC,
USA
BUM-RAK CHOI • Cardiovascular Research Center, Cardiovascular Institute, Rhode Island
Hospital and Alpert Medical School of Brown University, Providence, RI, USA
KAREN L. CHRISTMAN • Sanford Consortium for Regenerative Medicine, Department of
Bioengineering, University of California San Diego, La Jolla, CA, USA
BRIAN D. COFFIN • Department of Materials Science and Engineering, Carnegie Mellon
University, Pittsburgh, PA, USA
KAREEN L. K. COULOMBE • School of Engineering, Center for Biomedical Engineering,
Brown University, Providence, RI, USA
NATHAN CUNNINGHAM • Stanford Cardiovascular Institute, Stanford University, Stanford,
CA, USA; Division of Cardiovascular Medicine, Department of Medicine, Stanford
University, Stanford, CA, USA
MICHAEL E. DAVIS • Wallace H. Coulter Department of Biomedical Engineering, Georgia
Institute of Technology and Emory University School of Medicine, Atlanta, GA, USA;
Division of Cardiology, Department of Medicine, Emory University School of Medicine,
Atlanta, GA, USA; Children’s Heart Research and Outcomes (HeRO) Center, Children’s
Healthcare of Atlanta and Emory University, Atlanta, GA, USA
AMY L. DE JONGH CURRY • Department of Biomedical Engineering, University of Memphis,
Memphis, TN, USA
SOPHIA DELUCA • Department of Cell Biology, Duke University, Durham, NC, USA

xi
xii Contributors

ALEXANDRA EDER • Department of Experimental Pharmacology and Toxicology, University

Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany; DZHK (German
Centre for Cardiovascular Research), Hamburg/Kiel/Lübeck, Germany
EMILIA ENTCHEVA • Department of Biomedical Engineering, School of Engineering and
Applied Science, The George Washington University, Washington, DC, USA
THOMAS ESCHENHAGEN • Department of Experimental Pharmacology and Toxicology,
University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany; DZHK
(German Centre for Cardiovascular Research), Hamburg/Kiel/Lübeck, Germany
ADAM W. FEINBERG • Department of Materials Science and Engineering, Carnegie Mellon
University, Pittsburgh, PA, USA; Department of Biomedical Engineering, Carnegie
Mellon University, Pittsburgh, PA, USA
ALEX J. GOLDSTEIN • Institute for Stem Cell and Regenerative Medicine, University of
Washington, Seattle, WA, USA; Department of Materials Science and Engineering,
University of Washington, Seattle, WA, USA; Department of Laboratory Medicine &
Pathology, University of Washington, Seattle, WA, USA
JULIE L. HAN • Department of Biomedical Engineering, School of Engineering and Applied
Science, The George Washington University, Washington, DC, USA
TY HIGASHI • Institute for Stem Cell and Regenerative Medicine, University of Washington,
Seattle, WA, USA; Department of Mechanical Engineering, University of Washington,
Seattle, WA, USA
YI HONG • Department of Bioengineering, University of Texas at Arlington, Arlington, TX,
USA
ANDREW R. HUDSON • Department of Biomedical Engineering, Carnegie Mellon University,
Pittsburgh, PA, USA
NATHANIEL HUEBSCH • Department of Biomedical Engineering, Washington University in
St. Louis, St. Louis, MO, USA; NSF Science and Technology Center for Engineering
Mechanobiology, Washington University in St. Louis, St. Louis, MO, USA; Center for
Cardiovascular Research, Center for Regenerative Medicine, Center for Investigation of
Membrane Excitability Diseases, Washington University in St. Louis, St. Louis, MO, USA
JERVAUGHN D. HUNTER • Sanford Consortium for Regenerative Medicine, Department of
Bioengineering, University of California San Diego, La Jolla, CA, USA
TODD D. JOHNSON • Sanford Consortium for Regenerative Medicine, Department of
Bioengineering, University of California San Diego, La Jolla, CA, USA
CELINDA M. KOFRON • School of Engineering, Center for Biomedical Engineering, Brown
University, Providence, RI, USA
JULIA KRAUSE • Department of Experimental Pharmacology and Toxicology, University
Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany; DZHK (German
Centre for Cardiovascular Research), Hamburg/Kiel/Lübeck, Germany
RAYMOND K. KUDEJ • Department of Clinical Sciences, Tufts University School of Veterinary
Medicine, North Grafton, MA, USA
MICHAEL A. LAFLAMME • McEwen Stem Cell Institute, University Health Network, Toronto,
ON, Canada; Peter Munk Cardiac Centre, University Health Network, Toronto, ON,
Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto,
Toronto, ON, Canada
ANDREW LEE • Department of Biomedical Engineering, Carnegie Mellon University,
Pittsburgh, PA, USA
 Contributors xiii

MARTA LEMME • Department of Experimental Pharmacology and Toxicology, University

Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany; DZHK (German
Centre for Cardiovascular Research), Hamburg/Kiel/Lübeck, Germany
JUN LIAO • Department of Bioengineering, University of Texas at Arlington, Arlington, TX,
USA
WEIZHEN LI • Department of Biomedical Engineering, School of Engineering and Applied
Science, The George Washington University, Washington, DC, USA
AHMED I. MAHMOUD • Department of Cell and Regenerative Biology, University of
Wisconsin-Madison School of Medicine and Public Health, Madison, WI, USA; University
of Wisconsin-Madison School of Medicine and Public Health, Madison, WI, USA
INGRA MANNHARDT • Department of Experimental Pharmacology and Toxicology, University
Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany; DZHK (German
Centre for Cardiovascular Research), Hamburg/Kiel/Lübeck, Germany
ORIANE B. MATTHYS • UC Berkeley-UC San Francisco Graduate Program in
Bioengineering, San Francisco, CA, USA; Gladstone Institutes, San Francisco, CA, USA
MEGAN L. MCCAIN • Laboratory for Living Systems Engineering, Department of Biomedical
Engineering, USC Viterbi School of Engineering, University of Southern California, Los
Angeles, CA, USA; Department of Stem Cell Biology and Regenerative Medicine, Keck
School of Medicine of USC, University of Southern California, Los Angeles, CA, USA
TODD C. MCDEVITT • Gladstone Institutes, San Francisco, CA, USA; Department of
Bioengineering and Therapeutic Sciences, University of California, San Francisco, CA,
USA
SARAH MCLAUGHLIN • BioEngineering and Therapeutic Solutions (BEaTS), Division of
Cardiac Surgery, University of Ottawa Heart Institute, Ottawa, ON, Canada;
Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON,
Canada
TIM MEYER • Institute of Pharmacology and Toxicology, University Medical Center
Göttingen, Göttingen, Germany; DZHK (German Center for Cardiovascular Research),
Partner Site Göttingen, Göttingen, Germany
TERUO OKANO • Institute of Advanced Biomedical Engineering and Science, Tokyo Women’s
Medical University, Tokyo, Japan; Center for Advanced Medical and Life Science, Tokyo
Women’s Medical University, Tokyo, Japan; Cell Sheet Tissue Engineering Center
(CSTEC), School of Medicine and College of Pharmacy, Department of Pharmaceutics and
Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT, USA
ANDREW P. PETERSEN • Laboratory for Living Systems Engineering, Department of
Biomedical Engineering, USC Viterbi School of Engineering, University of Southern
California, Los Angeles, CA, USA
ELIZABETH C. PORTER • Cellular, Molecular and Developmental Biology Program, Graduate
School of Biomedical Sciences, Tufts University, Boston, MA, USA
BEIPING QIANG • McEwen Stem Cell Institute, University Health Network, Toronto, ON,
Canada
MILICA RADISIC • Institute of Biomedical Engineering, University of Toronto, Toronto, ON,
Canada; Department of Chemical Engineering and Applied Chemistry, University of
Toronto, Toronto, ON, Canada; Toronto General Research Institute, University Health
Network, Toronto, ON, Canada
xiv Contributors

ROCCO ROMAGNUOLO • McEwen Stem Cell Institute, University Health Network, Toronto,
ON, Canada
PIERRE-LUC SATIN • Institute of Pharmacology and Toxicology, University Medical Center
Göttingen, Göttingen, Germany; DZHK (German Center for Cardiovascular Research),
Partner Site Göttingen, Göttingen, Germany
HIDEKAZU SEKINE • Institute of Advanced Biomedical Engineering and Science, Tokyo
Women’s Medical University, Tokyo, Japan; Center for Advanced Medical and Life Science,
Tokyo Women’s Medical University, Tokyo, Japan; Cell Sheet Tissue Engineering Center
(CSTEC), School of Medicine and College of Pharmacy, Department of Pharmaceutics and
Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT, USA
MICKEY SHAH • Department of Biomedical Engineering, The University of Akron, Akron,
OH, USA
PREETY SHAKYA • Wallace H. Coulter Department of Biomedical Engineering, Georgia
Institute of Technology and Emory University School of Medicine, Atlanta, GA, USA
DANIEL W. SIMMONS • Department of Biomedical Engineering, Washington University in St.
Louis, St. Louis, MO, USA; NSF Science and Technology Center for Engineering
Mechanobiology, Washington University in St. Louis, St. Louis, MO, USA
JACOB SMITH • Department of Chemical Engineering and Applied Chemistry, University of
Toronto, Toronto, ON, Canada
DAVID SMYTH • Cardiac Function Laboratory, University of Ottawa Heart Institute,
Ottawa, ON, Canada
NATHAN J. SNIADECKI • Institute for Stem Cell and Regenerative Medicine, University of
Washington, Seattle, WA, USA; Department of Bioengineering, University of Washington,
Seattle, WA, USA; Department of Mechanical Engineering, University of Washington,
Seattle, WA, USA; Department of Laboratory Medicine & Pathology, University of
Washington, Seattle, WA, USA
JUSTUS STENZIG • Department of Experimental Pharmacology and Toxicology, University
Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany; DZHK (German
Centre for Cardiovascular Research), Hamburg/Kiel/Lübeck, Germany
WHITNEY L. STOPPEL • Department of Biomedical Engineering, Tufts University, Medford,
MA, USA; Department of Chemical Engineering, University of Florida, Gainesville, FL,
USA
ERIK J. SUURONEN • BioEngineering and Therapeutic Solutions (BEaTS), Division of
Cardiac Surgery, University of Ottawa Heart Institute, Ottawa, ON, Canada;
Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON,
Canada
MALTE TIBURCY • Institute of Pharmacology and Toxicology, University Medical Center
Göttingen, Göttingen, Germany; DZHK (German Center for Cardiovascular Research),
Partner Site Göttingen, Göttingen, Germany
BA€ RBEL ULMER • Department of Experimental Pharmacology and Toxicology, University
Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany; DZHK (German
Centre for Cardiovascular Research), Hamburg/Kiel/Lübeck, Germany
BO WANG • Joint Department of Biomedical Engineering, Marquette University and the
Medical College of Wisconsin, Milwaukee, WI, USA
ERIKA YAN WANG • Institute of Biomedical Engineering, University of Toronto, Toronto, ON,
Canada
LAKIESHA N. WILLIAMS • Department of Biomedical Engineering, University of Florida,
Gainesville, FL, USA
 Contributors xv

JOSEPH C. WU • Stanford Cardiovascular Institute, Stanford University, Stanford, CA,

USA; Division of Cardiovascular Medicine, Department of Medicine, Stanford University,
Stanford, CA, USA; Department of Radiology, Stanford University School of Medicine,
Stanford, CA, USA
FANNY WULKAN • McEwen Stem Cell Institute, University Health Network, Toronto, ON,
Canada
GE ZHANG • Department of Biomedical Engineering, The University of Akron, Akron, OH,
USA
WOLFRAM-HUBERTUS ZIMMERMANN • Institute of Pharmacology and Toxicology, University
Medical Center Göttingen, Göttingen, Germany; DZHK (German Center for
Cardiovascular Research), Partner Site Göttingen, Göttingen, Germany; Cluster of
Excellence “Multiscale Bioimaging: From Molecular Machines to Networks of Excitable
Cells” (MBExC), University of Göttingen, Göttingen, Germany; Center for
Neurodegenerative Diseases (DZNE), Göttingen, Germany; Fraunhofer Institute for
Translational Medicine and Pharmacology (ITMP), Göttingen, Germany
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