Human Organotypic Cultured Cardiac Slices: New Platform For High Throughput Preclinical Human Trials

Kang, Chaoyi; Qiao, Yun; Li, G; Baechle, K; Camelliti, Patrizia; Rentschler, Stacey; Efimov, Igor R.

doi:10.1038/srep28798

Cited by 90 publications

(99 citation statements)

References 44 publications

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“…; Kang et al . ). This report is the first to present a detailed protocol for successful preparation of transverse ventricular slices for transmural and regional profiling of V m and CaT with high resolution optical mapping.…”

Section: Discussionmentioning

confidence: 97%

“…; Kang et al . ). Furthermore, prior studies have been limited to the analysis of sub‐epicardial (Wang et al .…”

Section: Introductionmentioning

confidence: 97%

“…) or sub‐endocardial tangentially cut slices (Kang et al . ), or a few transmurally cut slices (Bussek et al . ), an approach inadequate for accurate capture of regional and transmural cardiac electrophysiological properties.…”

Section: Introductionmentioning

confidence: 99%

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Transverse cardiac slicing and optical imaging for analysis of transmural gradients in membrane potential and Ca²⁺ transients in murine heart

Wen

Gandhi

Capel

et al. 2018

The Journal of Physiology

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Key points A robust cardiac slicing approach was developed for optical mapping of transmural gradients in transmembrane potential (V m) and intracellular Ca2+ transient (CaT) of murine heart.Significant transmural gradients in V m and CaT were observed in the left ventricle.Frequency‐dependent action potentials and CaT alternans were observed in all ventricular regions with rapid pacing, with significantly greater incidence in the endocardium than epicardium.The observations demonstrate the feasibility of our new approach to cardiac slicing for systematic analysis of intrinsic transmural and regional gradients in V m and CaT. AbstractTransmural and regional gradients in membrane potential and Ca2+ transient in the murine heart are largely unexplored. Here, we developed and validated a robust approach which combines transverse ultra‐thin cardiac slices and high resolution optical mapping to enable systematic analysis of transmural and regional gradients in transmembrane potential (V m) and intracellular Ca2+ transient (CaT) across the entire murine ventricles. The voltage dye RH237 or Ca2+ dye Rhod‐2 AM were loaded through the coronary circulation using a Langendorff perfusion system. Short‐axis slices (300 μm thick) were prepared from the entire ventricles (from the apex to the base) by using a high‐precision vibratome. Action potentials (APs) and CaTs were recorded with optical mapping during steady‐state baseline and rapid pacing. Significant transmural gradients in V m and CaT were observed in the left ventricle, with longer AP duration (APD50 and APD75) and CaT duration (CaTD50 and CaTD75) in the endocardium compared with that in the epicardium. No significant regional gradients were observed along the apico‐basal axis of the left ventricle. Interventricular gradients were detected with significantly shorter APD50, APD75 and CaTD50 in the right ventricle compared with left ventricle and ventricular septum. During rapid pacing, AP and CaT alternans were observed in most ventricular regions, with significantly greater incidence in the endocardium in comparison with epicardium. In conclusion, these observations demonstrate the feasibility of our new approach to cardiac slicing for systematic analysis of intrinsic transmural and regional gradients in V m and CaT in murine ventricular tissue.

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Section: Discussionmentioning

confidence: 97%

“…; Kang et al . ). Furthermore, prior studies have been limited to the analysis of sub‐epicardial (Wang et al .…”

Section: Introductionmentioning

confidence: 97%

See 1 more Smart Citation

Transverse cardiac slicing and optical imaging for analysis of transmural gradients in membrane potential and Ca²⁺ transients in murine heart

Wen

Gandhi

Capel

et al. 2018

The Journal of Physiology

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“…23 Our final experimental platform was adult human ventricular tissue slices which may be useful as physiologicallyrelevant model systems for electrophysiology studies. 24…”

Section: Introductionmentioning

confidence: 99%

Adeno-associated virus mediated gene delivery: Implications for scalablein vitroandin vivocardiac optogenetic models

Ambrosi

Sadananda

Klimas

et al. 2017

Preprint

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Aims: Adeno-associated viruses (AAVs) provide advantages in long-term, cardiac-specific gene expression. However, AAV serotype specificity data is lacking in cardiac models relevant to optogenetics. We aimed to identify the optimal AAV serotype (1, 6, or 9) in pursuit of scalable rodent and human models for cardiac optogenetics and elucidate the mechanism of virus uptake. Methods:In vitro syncytia of primary neonatal rat ventricular cardiomyocytes (NRVMs) and human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were infected with AAVs 1, 6, and 9 containing the transgene for eGFP or channelrhodopsin-2 (ChR2) fused to mCherry. In vivo adult rats were intravenously injected with AAV1 and 9 containing ChR2-mCherry.Results: Transgene expression profiles of rat and human cells in vitro revealed that AAV1 and 6 significantly outperformed AAV9. In contrast, systemic delivery of AAV9 in adult rat hearts yielded significantly higher levels of ChR2-mCherry expression and optogenetic responsiveness. We tracked the mechanism of virus uptake to purported receptor-mediators for AAV1/6 (cell surface sialic acid) and AAV9 (37/67kDa laminin receptor, LamR). In vitro desialylation of NRVMs and hiPSC-CMs with neuraminidase significantly decreased AAV1,6-mediated gene expression, but interestingly, desialylation of hiPSC-CMs increased AAV9-mediated expression. In fact, only very high viral doses of AAV9-ChR2-mCherry, combined with neuraminidase treatment yielded consistent optogenetic responsiveness in hiPSC-CMs. Differences between the in vitro and in vivo performance of AAV9 could be correlated to robust LamR expression in the adult and neonatal rat hearts, but no expression in vitro in cultured cells. The dynamic nature of LamR expression and its dependence on environmental factors was further corroborated in intact adult human ventricular tissue slices.

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“…However, electrophysiological data obtained using healthy human adult myocardium are rare and difficult to obtain. A recent demonstration of maintaining physiological state of thin slices of human donor hearts for electrophysiological assessments (Kang et al 2016) is one of promising technologies to determine such parameters.…”

Section: Discussionmentioning

confidence: 99%

Computational modeling for cardiac safety pharmacology analysis: Contribution of fibroblasts

Gao

Engel

Carlson

et al. 2017

Journal of Pharmacological and Toxicological Methods

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Introduction Drug-induced proarrhythmic potential is an important regulatory criterion in safety pharmacology. The application of in silico approaches to predict proarrhythmic potential of new compounds is under consideration as part of future guidelines. Current approaches simulate electrophysiology of a single human adult ventricular cardiomyocyte. However, drug-induced proarrhythmic potential can be different when cardiomyocytes are surrounded by non-muscle cells. Incorporating fibroblasts in models of myocardium is important particularly for predicting a drugs cardiac liability in the aging population – a growing population who take more medications and exhibit increased cardiac fibrosis. In this study, we used computational models to investigate the effects of fibroblast coupling on the electrophysiology and response to drugs of cardiomyocytes. Methods A computational model of cardiomyocyte electrophysiology and ion handling (O’Hara et al. 2011) is coupled to a passive model of fibroblast electrophysiology to test the effects of dofetilide block on the rapid delayed rectifier K+ channel. Results are compared to model results without fibroblast coupling to see how fibroblasts affect cardiomyocyte action potential duration at 90% repolarization (APD90) and propensity for early after depolarization (EAD). Results Simulation results show an increase in cardiomyocyte APD90 with increasing concentration of three drugs that affect cardiac function: dofetilide, vardenafil and nebivolol, when no fibroblasts are coupled to the cardiomyocyte. Coupling fibroblasts to cardiomyocytes markedly shortens APD90. Moreover, increasing the number of fibroblasts can augment the shortening effect. Discussion Coupling cardiomyocytes and fibroblasts are predicted to decrease proarrhythmic susceptibility under dofetilide, vardenafil and nebivolol block. However, this result is sensitive to parameters which define the electrophysiological function of the fibroblast. Fibroblast membrane capacitance and conductance (CFB and GFB) have less of an effect on APD90 than the fibroblast resting membrane potential (EFB). This study suggests that in both theoretical models and experimental tissue constructs that represent cardiac tissue, both cardiomyocytes and nonmuscle cells should be considered when testing cardiac pharmacological agents.

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Human Organotypic Cultured Cardiac Slices: New Platform For High Throughput Preclinical Human Trials

Cited by 90 publications

References 44 publications

Transverse cardiac slicing and optical imaging for analysis of transmural gradients in membrane potential and Ca²⁺ transients in murine heart

Transverse cardiac slicing and optical imaging for analysis of transmural gradients in membrane potential and Ca²⁺ transients in murine heart

Adeno-associated virus mediated gene delivery: Implications for scalablein vitroandin vivocardiac optogenetic models

Computational modeling for cardiac safety pharmacology analysis: Contribution of fibroblasts

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Human Organotypic Cultured Cardiac Slices: New Platform For High Throughput Preclinical Human Trials

Cited by 90 publications

References 44 publications

Transverse cardiac slicing and optical imaging for analysis of transmural gradients in membrane potential and Ca2+ transients in murine heart

Transverse cardiac slicing and optical imaging for analysis of transmural gradients in membrane potential and Ca2+ transients in murine heart

Adeno-associated virus mediated gene delivery: Implications for scalablein vitroandin vivocardiac optogenetic models

Computational modeling for cardiac safety pharmacology analysis: Contribution of fibroblasts

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Transverse cardiac slicing and optical imaging for analysis of transmural gradients in membrane potential and Ca²⁺ transients in murine heart

Transverse cardiac slicing and optical imaging for analysis of transmural gradients in membrane potential and Ca²⁺ transients in murine heart