Automated Video-Based Analysis of Contractility and Calcium Flux in Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes Cultured over Different Spatial Scales

Huebsch, Nathaniel; Loskill, Peter; Mandegar, Mohammad A.; Marks, Natalie C.; Sheehan, Alice S.; Ma, Zhen; Mathur, Anurag; Nguyen, Trieu N.; Yoo, Jennie; Judge, Luke M.; Spencer, C. Ian; Chukka, Anand C.; Russell, Caitlin R.; So, Po Lin; Conklin, Bruce R.; Healy, Kevin E.

doi:10.1089/ten.tec.2014.0283

Cited by 262 publications

(276 citation statements)

References 28 publications

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“…For chemical and genetically encoded Ca 2 + ‐sensitive dyes, the situation is easier, mainly due to the slower kinetics of Ca 2 + fluxes. These probes have already been widely used for high‐throughput screening in both disease modelling and drug development with encouraging results (Kaestner et al , 2014; Huebsch et al , 2015; Lu et al , 2015; Rast et al , 2015; Dempsey et al , 2016; Klimas et al , 2016). Ca 2 + ‐handling also affects cell contractility, which can also be measured, as recent examples with sophisticated optical mapping systems show (Herron et al , 2012; Hayakawa et al , 2014; Maddah et al , 2015).…”

Section: Assays and Readoutsmentioning

confidence: 99%

Integrating cardiomyocytes from human pluripotent stem cells in safety pharmacology: has the time come?

Sala

Bellin

Mummery

2016

British J Pharmacology

107

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Cardiotoxicity is a severe side effect of drugs that induce structural or electrophysiological changes in heart muscle cells. As a result, the heart undergoes failure and potentially lethal arrhythmias. It is still a major reason for drug failure in preclinical and clinical phases of drug discovery. Current methods for predicting cardiotoxicity are based on guidelines that combine electrophysiological analysis of cell lines expressing ion channels ectopically in vitro with animal models and clinical trials. Although no new cases of drugs linked to lethal arrhythmias have been reported since the introduction of these guidelines in 2005, their limited predictive power likely means that potentially valuable drugs may not reach clinical practice. Human pluripotent stem cell‐derived cardiomyocytes (hPSC‐CMs) are now emerging as potentially more predictive alternatives, particularly for the early phases of preclinical research. However, these cells are phenotypically immature and culture and assay methods not standardized, which could be a hurdle to the development of predictive computational models and their implementation into the drug discovery pipeline, in contrast to the ambitions of the comprehensive pro‐arrhythmia in vitro assay (CiPA) initiative. Here, we review present and future preclinical cardiotoxicity screening and suggest possible hPSC‐CM‐based strategies that may help to move the field forward. Coordinated efforts by basic scientists, companies and hPSC banks to standardize experimental conditions for generating reliable and reproducible safety indices will be helpful not only for cardiotoxicity prediction but also for precision medicine.Linked ArticlesThis article is part of a themed section on New Insights into Cardiotoxicity Caused by Chemotherapeutic Agents. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v174.21/issuetoc

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Section: Assays and Readoutsmentioning

confidence: 99%

Integrating cardiomyocytes from human pluripotent stem cells in safety pharmacology: has the time come?

Sala

Bellin

Mummery

2016

British J Pharmacology

107

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“…The cardiac microchambers can be characterized through immunocytochemistry with cardiomyocyte markers (cardiac troponin T, sarcomeric α-actinin, and myosin heavy chain). The physiological contraction of the beating chambers can be further quantitatively characterized using open-source in-house developed software 41 . A schematic outlining the entire procedure is illustrated in Fig.…”

Section: Introductionmentioning

confidence: 99%

Generation of spatial-patterned early-developing cardiac organoids using human pluripotent stem cells

et al. 2018

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The discovery of human induced pluripotent stem cells (hiPSCs) has provided an unprecedented opportunity to study tissue morphogenesis and organ development as “organogenesis-in-a-dish”. Current approaches of cardiac organoid engineering rely on either direct cardiac differentiation from embryoid bodies or generation of aligned cardiac tissues from pre-differentiated cardiomyocytes from monolayer hiPSCs. To experimentally model early cardiac organogenesis in vitro, our protocol combines biomaterials-based cell patterning with stem cell organoid engineering. Three-dimensional cardiac microchambers are created from two-dimensional hiPSC colonies, which approximates an early developing heart with distinct spatial organization and self-assembly. With proper training on photolithography microfabrication, maintenance of human pluripotent stem cells, and cardiac differentiation, a graduate student with guidance will likely be able to carry out this experimental protocol, which requires approximately three weeks. We envisage that this in vitro model of human early heart development could serve as an embryotoxicity screening assay in drug discovery, regulation, and prescription for healthy fetal development. Currently, this protocol has yet to be implemented to assess the cardiac microchamber formation from the hiPSC lines with inherited disease, which can potentially be used to study the disease mechanisms of cardiac malformations at early stage of embryogenesis.

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“…A much more comprehensive battery of techniques is already in regular use in the pharmaceutical industry, including genomics, transcriptomics, and proteomics. In recent MPS studies, we are seeing functional measurements such as contractility, calcium signaling, and electrophysiological responses of cardiac and skeletal muscle, 37,[69][70][71][72][73][74][75] electrical resistance and molecular permeability of barriers, [76][77][78][79][80][81][82] and metabolomic responses to inflammatory challenges. 78 In this issue, the Parker group uses a variety of measures, including gene expression profiling, to quantify factors that affect maturation of neonatal rat cardiomyocytes into a more mature phenotype.…”

Section: Fitting Into the "Grand Scheme"mentioning

confidence: 99%

Fitting tissue chips and microphysiological systems into the grand scheme of medicine, biology, pharmacology, and toxicology

Watson

Hunziker

Wikswo

2017

Exp Biol Med (Maywood)

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Impact statementMicrophysiological systems (MPS), which include engineered organoids and both individual and coupled organs-on-chips and tissue chips, are a rapidly growing topic of research that addresses the known limitations of conventional cellular monoculture on flat plastic -a wellperfected set of techniques that produces reliable, statistically significant results that may not adequately represent human biology and disease. As reviewed in this article and the others in this thematic issue, MPS research has made notable progress in the past three years in both cell sourcing and characterization. As the field matures, currently identified challenges are being addressed, and new ones are being recognized. Building upon investments by the Defense Advanced Research Projects Agency, National Institutes of Health, Food and Drug Administration, Defense Threat Reduction Agency, and Environmental Protection Agency of more than $200 million since 2012 and sizable corporate spending, academic and commercial players in the MPS community are demonstrating their ability to meet the translational challenges required to apply MPS technologies to accelerate drug development and advance toxicology. AbstractMicrophysiological systems (MPS), which include engineered organoids (EOs), single organ/tissue chips (TCs), and multiple organs interconnected to create miniature in vitro models of human physiological systems, are rapidly becoming effective tools for drug development and the mechanistic understanding of tissue physiology and pathophysiology. The second MPS thematic issue of Experimental Biology and Medicine comprises 15 articles by scientists and engineers from the National Institutes of Health, the IQ Consortium, the Food and Drug Administration, and Environmental Protection Agency, an MPS company, and academia. Topics include the progress, challenges, and future of organs-on-chips, dissemination of TCs into Pharma, children's health protection, liver zonation, liver chips and their coupling to interconnected systems, gastrointestinal MPS, maturation of immature cardiomyocytes in a heart-on-a-chip, coculture of multiple cell types in a human skin construct, use of synthetic hydrogels to create EOs that form neural tissue models, the blood-brain barrier-on-a-chip, MPS models of coupled female reproductive organs, coupling MPS devices to create a body-on-a-chip, and the use of a microformulator to recapitulate endocrine circadian rhythms. While MPS hardware has been relatively stable since the last MPS thematic issue, there have been significant advances in cell sourcing, with increased reliance on human-induced pluripotent stem cells, and in characterization of the genetic and functional cell state in MPS bioreactors. There is growing appreciation of the need to minimize perfusate-to-cell-volume ratios and respect physiological scaling of coupled TCs. Questions asked by drug developers are followed by an analysis of the potential value, costs, and needs of Pharma. Of highest value and lowest switching costs may be the d...

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Automated Video-Based Analysis of Contractility and Calcium Flux in Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes Cultured over Different Spatial Scales

Cited by 262 publications

References 28 publications

Integrating cardiomyocytes from human pluripotent stem cells in safety pharmacology: has the time come?

Integrating cardiomyocytes from human pluripotent stem cells in safety pharmacology: has the time come?

Generation of spatial-patterned early-developing cardiac organoids using human pluripotent stem cells

Fitting tissue chips and microphysiological systems into the grand scheme of medicine, biology, pharmacology, and toxicology

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