Passive Stretch Induces Structural and Functional Maturation of Engineered Heart Muscle as Predicted by Computational Modeling

Published on Feb 1, 2018in Stem Cells5.614
· DOI :10.1002/stem.2732
Oscar J. Abilez24
Estimated H-index: 24
Evangeline Tzatzalos4
Estimated H-index: 4
(Stanford University)
+ 18 AuthorsJoseph C. Wu91
Estimated H-index: 91
Background: The ability to differentiate human pluripotent stem cells (hPSCs) into cardiomyocytes (CMs) makes them an attractive source for repairing injured myocardium, disease modeling, and drug testing. Although current differentiation protocols yield hPSC-CMs to >90% efficiency, hPSC-CMs exhibit immature characteristics. With the goal of overcoming this limitation, we tested the effects of varying passive stretch on engineered heart muscle (EHM) structural and functional maturation, guided by computational modeling. Methods and Results: Human embryonic stem cells (hESCs, H7 line) or human induced pluripotent stem cells (hiPSCs, IMR-90 line) were differentiated to human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) in vitro using a small molecule based protocol. hPSC-CMs were characterized by troponin+ flow cytometry as well as electrophysiological measurements. Afterwards, 1.2 x 106 hPSC-CMs were mixed with 0.4 x 106 human fibroblasts (IMR-90 line) (3:1 ratio) and Type-I collagen. The blend was cast into custom-made 12-mm long polydimethylsiloxane (PDMS) reservoirs to vary nominal passive stretch of EHMs to 5, 7, or 9 mm. EHM characteristics were monitored for up to 50 days, with EHMs having a passive stretch of 7 mm giving the most consistent formation. Based on our initial macroscopic observations of EHM formation, we created a computational model that predicts the stress distribution throughout EHMs, which is a function of cellular composition, cellular ratio, and geometry. Based on this predictive modeling, we show cell alignment by immunohistochemistry and coordinated calcium waves by calcium imaging. Furthermore, coordinated calcium waves and mechanical contractions were apparent throughout entire EHMs. The stiffness and active forces of hPSC-derived EHMs are comparable to rat neonatal cardiomyocyte-derived EHMs. Three-dimensional EHMs display increased expression of mature cardiomyocyte genes including sarcomeric protein troponin-T, calcium and potassium ion channels, β-adrenergic receptors, and t-tubule protein caveolin-3. Conclusions: Passive stretch affects the structural and functional maturation of EHMs. Based on our predictive computational modeling, we show how to optimize cell alignment and calcium dynamics within EHMs. These findings provide a basis for the rational design of EHMs, which enables future scale-up productions for clinical use in cardiovascular tissue engineering. This article is protected by copyright. All rights reserved.
  • References (62)
  • Citations (18)
📖 Papers frequently viewed together
142 Citations
126 Citations
17 Authors (Sara S. Nunes, ..., Milica Radisic)
369 Citations
78% of Scinapse members use related papers. After signing in, all features are FREE.
#1Malte TiburcyH-Index: 18
#2James E. HudsonH-Index: 14
view all 31 authors...
Background:Advancing structural and functional maturation of stem cell–derived cardiomyocytes remains a key challenge for applications in disease modeling, drug screening, and heart repair. Here, w...
126 CitationsSource
#1Bin Wei (WSU: Wayne State University)H-Index: 2
#2J.-P. Jin (WSU: Wayne State University)H-Index: 35
Abstract Troponin T (TnT) is a central player in the calcium regulation of actin thin filament function and is essential for the contraction of striated muscles. Three homologous genes have evolved in vertebrates to encode three muscle type-specific TnT isoforms: TNNT1 for slow skeletal muscle TnT, TNNT2 for cardiac muscle TnT, and TNNT3 for fast skeletal muscle TnT. Alternative splicing and posttranslational modifications confer additional structural and functional variations of TnT during deve...
31 CitationsSource
#1Nathaniel Huebsch (UCSF: University of California, San Francisco)H-Index: 24
#2Peter Loskill (University of California, Berkeley)H-Index: 18
Last. Bruce R. ConklinH-Index: 59
view all 19 authors...
Tissue engineering approaches have the potential to increase the physiologic relevance of human iPS-derived cells, such as cardiomyocytes (iPS-CM). However, forming Engineered Heart Muscle (EHM) typically requires >1 million cells per tissue. Existing miniaturization strategies involve complex approaches not amenable to mass production, limiting the ability to use EHM for iPS-based disease modeling and drug screening. Micro-scale cardiospheres are easily produced, but do not facilitate assembly ...
76 CitationsSource
#1Gwanghyun Jung (Stanford University)H-Index: 6
#2Giovanni Fajardo (Stanford University)H-Index: 18
Last. Daniel Bernstein (Stanford University)H-Index: 50
view all 14 authors...
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are a powerful platform for uncovering disease mechanisms and assessing drugs for efficacy/toxicity. However, the accuracy with which hiPSC-CMs recapitulate the contractile and remodeling signaling of adult cardiomyocytes is not fully known. We used β-adrenergic receptor (β-AR) signaling as a prototype to determine the evolution of signaling component expression and function during hiPSC-CM maturation. In “early” hiPSC-CMs (l...
24 CitationsSource
#1Evangeline Tzatzalos (Stanford University)H-Index: 4
#2Oscar J. AbilezH-Index: 24
Last. Joseph C. WuH-Index: 91
view all 4 authors...
Engineered heart tissue has emerged as a personalized platform for drug screening. With the advent of induced pluripotent stem cell (iPSC) technology, patient-specific stem cells can be developed and expanded into an indefinite source of cells. Subsequent developments in cardiovascular biology have led to efficient differentiation of cardiomyocytes, the force-producing cells of the heart. iPSC-derived cardiomyocytes (iPSC-CMs) have provided potentially limitless quantities of well-characterized,...
63 CitationsSource
#1Christopher J. Kane (NIH: National Institutes of Health)H-Index: 58
#2Liam S. Couch (NIH: National Institutes of Health)H-Index: 3
Last. Cesare M. Terracciano (NIH: National Institutes of Health)H-Index: 34
view all 3 authors...
Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) hold enormous potential in many fields of cardiovascular research. Overcoming many of the limitations of their embryonic counterparts, the application of iPSC-CMs ranges from facilitating investigation of familial cardiac disease and pharmacological toxicity screening to personalized medicine and autologous cardiac cell therapies. The main factor preventing the full realization of this potential is the limited maturity of iPSC-CMs, ...
24 CitationsSource
#1Ioannis Karakikes (Stanford University)H-Index: 27
#2Mohamed Ameen (Stanford University)H-Index: 9
Last. Joseph C. Wu (Stanford University)H-Index: 91
view all 4 authors...
Disease models are essential for understanding cardiovascular disease pathogenesis and developing new therapeutics. The human induced pluripotent stem cell (iPSC) technology has generated significant enthusiasm for its potential application in basic and translational cardiac research. Patient-specific iPSC-derived cardiomyocytes offer an attractive experimental platform to model cardiovascular diseases, study the earliest stages of human development, accelerate predictive drug toxicology tests, ...
128 CitationsSource
#1Lei Ye (UMN: University of Minnesota)H-Index: 12
#2Ying Hua Chang (UW: University of Wisconsin-Madison)H-Index: 2
Last. Jianyi ZhangH-Index: 41
view all 20 authors...
Summary Human induced pluripotent stem cells (hiPSCs) hold promise for myocardial repair following injury, but preclinical studies in large animal models are required to determine optimal cell preparation and delivery strategies to maximize functional benefits and to evaluate safety. Here, we utilized a porcine model of acute myocardial infarction (MI) to investigate the functional impact of intramyocardial transplantation of hiPSC-derived cardiomyocytes, endothelial cells, and smooth muscle cel...
185 CitationsSource
#1Marc N. Hirt (UHH: University of Hamburg)H-Index: 12
#2Jasper Boeddinghaus (UHH: University of Hamburg)H-Index: 16
Last. EschenhagenThomas (UHH: University of Hamburg)H-Index: 68
view all 15 authors...
Abstract Spontaneously beating engineered heart tissue (EHT) represents an advanced in vitro model for drug testing and disease modeling, but cardiomyocytes in EHTs are less mature and generate lower forces than in the adult heart. We devised a novel pacing system integrated in a setup for videooptical recording of EHT contractile function over time and investigated whether sustained electrical field stimulation improved EHT properties. EHTs were generated from neonatal rat heart cells (rEHT, n ...
133 CitationsSource
#1Paul W. Burridge (Stanford University)H-Index: 25
#2Elena Matsa (Stanford University)H-Index: 20
Last. Joseph C. Wu (Stanford University)H-Index: 91
view all 15 authors...
A simple, robust, chemically defined method for generating cardiomyocytes from human pluripotent stem cells is described. It should enable the identification of conditions for maturation of these cells.
513 CitationsSource
Cited By18
#1Yuxuan Guo (Boston Children's Hospital)H-Index: 5
#2William T. Pu (Harvard University)H-Index: 59
Maturation is the last phase of heart development that prepares the organ for strong, efficient, and persistent pumping throughout the mammal's lifespan. This process is characterized by structural, gene expression, metabolic, and functional specializations in cardiomyocytes as the heart transits from fetal to adult states. Cardiomyocyte maturation gained increased attention recently due to the maturation defects in pluripotent stem cell-derived cardiomyocyte, its antagonistic effect on myocardi...
#2Tatsuya Anzai (Jichi Medical University)H-Index: 1
Last. Hideki UosakiH-Index: 14
view all 4 authors...
Cardiovascular diseases are the leading cause of death worldwide. Therefore, the discovery of induced pluripotent stem cells (iPSCs) and the subsequent generation of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) was a pivotal point in regenerative medicine and cardiovascular research. They constituted an appealing tool for replacing dead and dysfunctional cardiac tissue, screening cardiac drugs and toxins, and studying inherited cardiac diseases. The problem is that thes...
1 CitationsSource
#1Chenyan Wang (SU: Syracuse University)
#1Chenyan Wang (SU: Syracuse University)H-Index: 1
Last. Zhen Ma (SU: Syracuse University)H-Index: 13
view all 9 authors...
Cardiac tissues are able to adjust their contractile behavior to adapt to the local mechanical environment. Nonuniformity of the native tissue mechanical properties contributes to the development of heart dysfunctions, yet the current in vitro cardiac tissue models often fail to recapitulate the mechanical nonuniformity. To address this issue, a 3D cardiac microtissue model is developed with engineered mechanical nonuniformity, enabled by 3D-printed hybrid matrices composed of fibers with differ...
#1Nicole Silbernagel (Heidelberg University)
#2Arlene Körner (Heidelberg University)H-Index: 1
Last. Nina D. Ullrich (KIT: Karlsruhe Institute of Technology)H-Index: 4
view all 11 authors...
Abstract Cardiomyocytes derived from induced pluripotent stem cells (iPSC-CMs) represent the best cell source for cardiac regenerative purposes but retain an immature phenotype after differentiation with significant limitations compared to adult cardiomyocytes. Apart from an incomplete cardiomyocyte-specific structure and microarchitecture, cells show at the level of Ca2+ signaling only slow Ca2+ release and reuptake properties. Here, we investigated the effect of restructuring single iPSC-CMs i...
1 CitationsSource
#1Gary A. GintantH-Index: 2
#1G. A. GintantH-Index: 7
Last. Joseph C. WuH-Index: 91
view all 8 authors...
It is now well recognized that many lifesaving oncology drugs may adversely affect the heart and cardiovascular system, including causing irreversible cardiac injury that can result in reduced qual...
#1Alexandre J. S. Ribeiro (CDER: Center for Drug Evaluation and Research)
#2Brian D. Guth (NWU: North-West University)H-Index: 34
Last. Brian R. Berridge (NIH: National Institutes of Health)H-Index: 1
view all 14 authors...
Contractility of the myocardium engines the pumping function of the heart and is enabled by the collective contractile activity of its muscle cells: cardiomyocytes. The effects of drugs on the contractility of human cardiomyocytes in vitro can provide mechanistic insight that can support the prediction of clinical cardiac drug effects early in drug development. Cardiomyocytes differentiated from human induced pluripotent stem cells have high potential for overcoming the current limitations of co...
#1Mitch Biermann (UW: University of Wisconsin-Madison)H-Index: 2
#2Wenxuan Cai (UW: University of Wisconsin-Madison)H-Index: 9
Last. Timothy J. Kamp (UW: University of Wisconsin-Madison)H-Index: 48
view all 21 authors...
1 CitationsSource
#1Nima Momtahan (University of Texas at Austin)
#2Cody O. Crosby (University of Texas at Austin)H-Index: 2
Last. Janet Zoldan (University of Texas at Austin)H-Index: 10
view all 3 authors...
Recent advances in developmental biology and biomedical engineering have significantly improved the efficiency and purity of cardiomyocytes (CMs) generated from pluripotent stem cells (PSCs). Regardless of the protocol used to derive CMs, these cells exhibit hallmarks of functional immaturity. In this Opinion, we focus on reactive oxygen species (ROS), signaling molecules that can potentially modulate cardiac maturation. We outline how ROS impacts nearly every aspect associated with cardiac matu...
#1R. Madonna (University of Texas at Austin)H-Index: 8
#2Linda W. van Laake (UU: Utrecht University)H-Index: 25
Last. Joost P.G. Sluijter (UU: Utrecht University)H-Index: 41
view all 21 authors...
8 CitationsSource
#1F. Sahli Costabal (Stanford University)H-Index: 1
#2Jenny S. ChoyH-Index: 9
Last. Ellen Kuhl (Stanford University)H-Index: 49
view all 6 authors...
Abstract Dilated cardiomyopathy is a progressive irreversible disease associated with contractile dysfunction and heart failure. During dilated cardiomyopathy, elevated diastolic wall strains trigger mechanotransduction pathways that initiate the addition of sarcomeres in series and an overall increase in myocyte length. At the whole organ level, this results in a chronic dilation of the ventricles, an increase in end diastolic and end systolic volumes, and a decrease in ejection fraction. Howev...
5 CitationsSource