A Systemized Approach to Investigate Ca2+ Synchronization in Clusters of Human Induced Pluripotent Stem-Cell Derived Cardiomyocytes

Jones, Ann Rosalind; Edwards, David Hughes; Cummins, Michael; Williams, Alan J.; George, Christopher H.

doi:10.3389/fcell.2015.00089

Cited by 5 publications

(3 citation statements)

References 82 publications

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“…Differentiation protocols can result in the development of a heterogeneous population of cardiac myocytes, as evidenced by varied electrophysiological parameters[ 39 ], calcium handling[ 40 ] and gene expression profiles[ 40 ]. To address this, we sought to examine the potential effects of geometric heterogeneity on physiologic force production.…”

Section: Resultsmentioning

confidence: 99%

Investigation of human iPSC-derived cardiac myocyte functional maturation by single cell traction force microscopy

Wheelwright¹,

Win²,

Mikkila³

et al. 2018

PLoS ONE

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Recent advances have made it possible to readily derive cardiac myocytes from human induced pluripotent stem cells (hiPSC-CMs). HiPSC-CMs represent a valuable new experimental model for studying human cardiac muscle physiology and disease. Many laboratories have devoted substantial effort to examining the functional properties of isolated hiPSC-CMs, but to date, force production has not been adequately characterized. Here, we utilized traction force microscopy (TFM) with micro-patterning cell printing to investigate the maximum force production of isolated single hiPSC-CMs under varied culture and assay conditions. We examined the role of length of differentiation in culture and the effects of varied extracellular calcium concentration in the culture media on the maturation of hiPSC-CMs. Results show that hiPSC-CMs developing in culture for two weeks produced significantly less force than cells cultured from one to three months, with hiPSC-CMs cultured for three months resembling the cell morphology and function of neonatal rat ventricular myocytes in terms of size, dimensions, and force production. Furthermore, hiPSC-CMs cultured long term in conditions of physiologic calcium concentrations were larger and produced more force than hiPSC-CMs cultured in standard media with sub-physiological calcium. We also examined relationships between cell morphology, substrate stiffness and force production. Results showed a significant relationship between cell area and force. Implementing directed modifications of substrate stiffness, by varying stiffness from embryonic-like to adult myocardium-like, hiPSC-CMs produced maximal forces on substrates with a lower modulus and significantly less force when assayed on increasingly stiff adult myocardium-like substrates. Calculated strain energy measurements paralleled these findings. Collectively, these findings further establish single cell TFM as a valuable approach to illuminate the quantitative physiological maturation of force in hiPSC-CMs.

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Section: Resultsmentioning

confidence: 99%

Investigation of human iPSC-derived cardiac myocyte functional maturation by single cell traction force microscopy

Wheelwright¹,

Win²,

Mikkila³

et al. 2018

PLoS ONE

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“…A consequence may then include differential responses to drugs. Jones et al proposed Ca 2 + synchronization in aggregates of hiPSC as a reliable parameter to identify sources of variability related to culture conditions and maturation status of hiPSC‐CMs (Jones et al , 2015). …”

Section: Limitations In Applicability Of Hipsc‐cm To Large‐scale Drugmentioning

confidence: 99%

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

Sala

Bellin

Mummery

2016

British J Pharmacology

106

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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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“…new approaches to map to progression of sub-clinical cardiovascular disease [65,66]. Factors that contribute to towards variability and mosaicism in in vitro platforms are being elucidated and may also help to explain why developing the next generation of drugs requires more than an understanding of the underlying biology [35,67,68,69,70,71,72,73].…”

Section: Defining the Physiological Gamut And Systems Limitsmentioning

confidence: 99%

Introduction to biological complexity as a missing link in drug discovery

Gintant

George

2018

Expert Opinion on Drug Discovery

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Despite a burgeoning knowledge of the intricacies and mechanisms responsible for human disease, technological advances in medicinal chemistry, and more efficient assays used for drug screening, it remains difficult to discover novel and effective pharmacologic therapies. Areas covered: By reference to the primary literature and concepts emerging from academic and industrial drug screening landscapes, the authors propose that this disconnect arises from the inability to scale and integrate responses from simpler model systems to outcomes from more complex and human-based biological systems. Expert opinion: Further collaborative efforts combining target-based and phenotypic-based screening along with systems-based pharmacology and informatics will be necessary to harness the technological breakthroughs of today to derive the novel drug candidates of tomorrow. New questions must be asked of enabling technologies-while recognizing inherent limitations-in a way that moves drug development forward. Attempts to integrate mechanistic and observational information acquired across multiple scales frequently expose the gap between our knowledge and our understanding as the level of complexity increases. We hope that the thoughts and actionable items highlighted will help to inform the directed evolution of the drug discovery process.

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A Systemized Approach to Investigate Ca2+ Synchronization in Clusters of Human Induced Pluripotent Stem-Cell Derived Cardiomyocytes

Cited by 5 publications

References 82 publications

Investigation of human iPSC-derived cardiac myocyte functional maturation by single cell traction force microscopy

Investigation of human iPSC-derived cardiac myocyte functional maturation by single cell traction force microscopy

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

Introduction to biological complexity as a missing link in drug discovery

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