Macro‐micro imaging of cardiac–neural circuits in co‐cultures from normal and diseased hearts

Bub, Gil; Burton, Rebecca A.B.

doi:10.1113/jphysiol.2014.285460

Cited by 5 publications

(7 citation statements)

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“…Tissue culture models provide experimental access to model the intricate connections between the PNS and CVS at the cellular level, a degree of access that is not possible using more traditional in vivo and ex vivo techniques. Cardiac tissue monolayers are excitable media that serve as an intermediary between in silico and in vivo experimentation [ 77 , 105 , 106 ]. Combining engineered tissue models with cellular electrophysiology, optical actuation and optical characterization further enhances the experimental precision accessible to characterize neuro-cardiac physiology and pathophysiology.…”

Section: Discussion and Future Directionsmentioning

confidence: 99%

“…Structured patterning of myocytes and neurons offers a new model to address complex questions relating to neuro-cardiac behaviour in models of physiology and pathophysiology where tissue properties are directionally dependent. Engineered tissue constructs support complex behaviours and recapitulate structure-function relationships enabling precise measurement of macroscopic behaviour (such as spiral re-entrant waves, figure 2c) [27,70,[75][76][77]. The translational capacity of tissue culture models has improved with the design of anisotropic tissues as well as the use of human-induced pluripotent stem cells (hiPSCs) [54,72,73,78,79].…”

Section: In Vitro Approaches To Examine Neuro-cardiac Interactionsmentioning

confidence: 99%

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Combining tissue engineering and optical imaging approaches to explore interactions along the neuro-cardiac axis

et al. 2020

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Interactions along the neuro-cardiac axis are being explored with regard to their involvement in cardiac diseases, including catecholaminergic polymorphic ventricular tachycardia, hypertension, atrial fibrillation, long QT syndrome and sudden death in epilepsy. Interrogation of the pathophysiology and pathogenesis of neuro-cardiac diseases in animal models present challenges resulting from species differences, phenotypic variation, developmental effects and limited availability of data relevant at both the tissue and cellular level. By contrast, tissue-engineered models containing cardiomyocytes and peripheral sympathetic and parasympathetic neurons afford characterization of cellular- and tissue-level behaviours while maintaining precise control over developmental conditions, cellular genotype and phenotype. Such approaches are uniquely suited to long-term, high-throughput characterization using optical recording techniques with the potential for increased translational benefit compared to more established techniques. Furthermore, tissue-engineered constructs provide an intermediary between whole animal/tissue experiments and in silico models. This paper reviews the advantages of tissue engineering methods of multiple cell types and optical imaging techniques for the characterization of neuro-cardiac diseases.

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Section: Discussion and Future Directionsmentioning

confidence: 99%

Section: In Vitro Approaches To Examine Neuro-cardiac Interactionsmentioning

confidence: 99%

Combining tissue engineering and optical imaging approaches to explore interactions along the neuro-cardiac axis

et al. 2020

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“…Dr Rebecca Burton presented her recent work on developing a cell culture model of neurally mediated arrhythmogenesis and non-invasive optical imaging methods being pursued at Oxford. 86 , 87 Biological models with varying degrees of complexity have been developed to shed light on re-entrant arrhythmias and cardiac monolayers are one of the simplest models. The next stages of this research is to pursue remote monitoring of in vitro cell cultures that would increase experimental access, reduce the need to sacrifice additional animals, and spur the adoption by other laboratories working in allied research areas.…”

Section: Description Of the Workhopmentioning

confidence: 99%

Human-based approaches to pharmacology and cardiology: an interdisciplinary and intersectorial workshop

et al. 2015

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Both biomedical research and clinical practice rely on complex datasets for the physiological and genetic characterization of human hearts in health and disease. Given the complexity and variety of approaches and recordings, there is now growing recognition of the need to embed computational methods in cardiovascular medicine and science for analysis, integration and prediction. This paper describes a Workshop on Computational Cardiovascular Science that created an international, interdisciplinary and inter-sectorial forum to define the next steps for a human-based approach to disease supported by computational methodologies. The main ideas highlighted were (i) a shift towards human-based methodologies, spurred by advances in new in silico, in vivo, in vitro, and ex vivo techniques and the increasing acknowledgement of the limitations of animal models. (ii) Computational approaches complement, expand, bridge, and integrate in vitro, in vivo, and ex vivo experimental and clinical data and methods, and as such they are an integral part of human-based methodologies in pharmacology and medicine. (iii) The effective implementation of multi- and interdisciplinary approaches, teams, and training combining and integrating computational methods with experimental and clinical approaches across academia, industry, and healthcare settings is a priority. (iv) The human-based cross-disciplinary approach requires experts in specific methodologies and domains, who also have the capacity to communicate and collaborate across disciplines and cross-sector environments. (v) This new translational domain for human-based cardiology and pharmacology requires new partnerships supported financially and institutionally across sectors. Institutional, organizational, and social barriers must be identified, understood and overcome in each specific setting.

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“…Cardiac impulse formation and conduction are modulated by autonomic activity, and the autonomic nervous system plays an important role in the initiation and maintenance of arrhythmias in diseased hearts [1,2]. Sympathetic nerves release noradrenaline, which activates cardiac β-adrenergic receptors to modulate myocyte repolarisation and calcium handling via alterations of transmembrane currents and intracellular calcium homeostasis [3].…”

Section: Introductionmentioning

confidence: 99%

Optical interrogation of sympathetic neuronal effects on macroscopic cardiac monolayer dynamics

Burton

Tomek

Ambrosi

et al. 2019

Preprint

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Alterations in autonomic function are known to occur in cardiac conditions including sudden cardiac death. Cardiac stimulation via sympathetic neurons can potentially trigger arrhythmias. Dissecting direct neural-cardiac interactions at the cellular level is technically challenging and understudied due to the lack of experimental model systems and methodologies. Here we demonstrate the utility of optical interrogation of sympathetic neurons and their effects on macroscopic cardiac monolayer dynamics to address research targets such as the effects of adrenergic stimulation via the release of neurotransmitters, the effect of neuronal numbers on cardiac wave behaviour and the applicability of optogenetics in mechanistic in vitro studies. We combine photo-uncaging or optogenetic neural stimulation with imaging of cardiac monolayers to measure electrical activity in an automated fashion, illustrating the power and high throughput capability of such interrogations. The methods described highlight the challenges and benefits of co-cultures as experimental model systems. RESULTS:1] Stellate Sympathetic Neurons make contacts with cardiomyocytes: Scanning electron microscopy of sympathetic neurons growing in co-culture with cardiomyocytes in vitro shows connections between neurite extension and cardiac syncytium (Figure 1 A). The neuron bodies and extensions clearly make physical contact with myocytes (Figure 1 B, C, D, E, F, G, H, I, J). Immuno staining of co-cultures shows co-localisation of Synpasin and Beta2 receptors (Figure 1 K), sympathetic neurons show positive staining for Th (Figure 1 L), and fibronectin contamination in the co-cultures was assessed by staining with Vimentin (Figure 1 M), which showed low abundance. 2] Pattern formation is affected by the presence of neurons: Using dye-free imaging (Figure 2 A, B, C), we investigated how neuronal activation modulates cardiac patterns of activation in monolayer culture. We chose experimental conditions that spontaneously yield a wide variety of wavefront topologies within the imaging system's 16 x 16 mm field of view. Introduction of an additional cell type can potentially introduce heterogeneities that would impact wave front stability. Surprisingly, co-cultures displayed fewer wave breaks than their monoculture counterparts at similar plating densities (Figure 2 D). We broadly classified wave dynamics as simple (periodic target waves and single spiral wave reentry, Figure 2 D top row), or complex (single dominant spirals with additional irregular waves and multiple equally sized wavelets, Figure 2 D bottom row). While monocultures frequently displayed complex dynamics, co-cultures rarely displayed this behavior (Figure 2 E, P<0.05, chi-square test). Indeed, we observed wavelet reentry in only one of the co-culture preparations (6 isolations, 20 preparations). 3] Conduction velocity in spontaneously active co-cultures is faster than myocyte monocultures:We measured conduction velocity in unstimulated co-cultures (n=19) and myocyte monocultures (n=9). Figure 2 F s...

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Macro‐micro imaging of cardiac–neural circuits in co‐cultures from normal and diseased hearts

Cited by 5 publications

References 50 publications

Combining tissue engineering and optical imaging approaches to explore interactions along the neuro-cardiac axis

Combining tissue engineering and optical imaging approaches to explore interactions along the neuro-cardiac axis

Human-based approaches to pharmacology and cardiology: an interdisciplinary and intersectorial workshop

Optical interrogation of sympathetic neuronal effects on macroscopic cardiac monolayer dynamics

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