Organotypic slice culture from human adult ventricular myocardium

Background/Aims: Safety pharmacology requires novel model systems for the detection of cardiac side effects. Ranging from cell-based systems to model organisms, no model available to date reflects the complexity of the human heart and evokes the great need for improved and more affordable systems. Many drugs interact with hERG potassium channels and consequently cause life threatening ventricular arrhythmias, further highlighting the importance of suitable model systems. Methods: Spontaneously Contracting Cell aggregates (SCC) as a 3D in vitro heart-syncytium obtained from rainbow trout larvae represent a novel model system for cardiac safety pharmacology. SCCs can be harvested cost-effectively and kept in culture for several weeks while retaining their functionality and displaying contraction rates similar to the human heart. Results: Extracellular field potential recordings with multielectrode arrays revealed significant prolongation of field potential duration upon administration of common hERG potassium channel blockers. Infusion of 1 µM Dofetilide and 10 µM Terfenadine prolonged field potentials 10 fold and 2 fold, respectively. In addition, SCCs enabled analysis of autonomous contraction frequencies. Conclusion: Thus, SCCs represent a novel and low-cost cardiac model system of the human heart for application in safety pharmacology.

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“…Human heart slices provide higher data quality, but cannot be used in medium - or high - throughput [34]. …”

Section: Discussionmentioning

confidence: 99%

Electrophysiological Characterization of Spontaneously Contracting Cell Aggregates Obtained from Rainbow Trout Larvae with Multielectrode Arrays

Mehnert

Brandenburger

Grunow

2013

Cell Physiol Biochem

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“…Adenosine receptor subtype-specific effects were also observed in isolated CMs. Antagonization of the A 1 R efficiently impeded the antihypertrophic effect of cAMP, thus restoring To investigate native cardiac tissue for cAMP-dependent communication between cells, we devised a near-vivo setup based on intact slices of myocardium that maintain organotypic functions for several days (22). Slices from hearts of 12-week-old mice were taken into culture, and sensor fibroblasts were transferred onto them.…”

Section: And Stefan Engelhardtmentioning

confidence: 99%

“…Vital slices of myocardial tissue were prepared from 12-week-old mice according to an established protocol (22). Fresh slices were seeded with trypsinized sensor fibroblasts and were maintained in coculture for a further 12 hours under conditions providing an air-liquid interface (Millicell-CM, Millipore) (22).…”

Section: Signaling Between Myocardial Slices and Sensor Cfs Through Smentioning

confidence: 99%

“…CFs were then trypsinized and seeded directly on the slices (prepared and incubated as described in ref. 22). After 12 hours, slices with adherent CFs were transferred to the experimental chamber in buffer C (137 mM NaCl, 5.4 mM KCl, 2 mM CaCl 2 , 1 mM MgCl 2 , 10 mM HEPES, pH 7.3) and images were taken as described above.…”

Section: Signaling Between Myocardial Slices and Sensor Cfs Through Smentioning

confidence: 99%

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Cardiac myocyte–secreted cAMP exerts paracrine action via adenosine receptor activation

Sassi¹,

Ahles²,

Truong³

et al. 2014

J. Clin. Invest.

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International audienceAcute stimulation of cardiac β-adrenoceptors is crucial to increasing cardiac function under stress; however, sustained β-adrenergic stimulation has been implicated in pathological myocardial remodeling and heart failure. Here, we have demonstrated that export of cAMP from cardiac myocytes is an intrinsic cardioprotective mechanism in response to cardiac stress. We report that infusion of cAMP into mice averted myocardial hypertrophy and fibrosis in a disease model of cardiac pressure overload. The protective effect of exogenous cAMP required adenosine receptor signaling. This observation led to the identification of a potent paracrine mechanism that is dependent on secreted cAMP. Specifically, FRET-based imaging of cAMP formation in primary cells and in myocardial tissue from murine hearts revealed that cardiomyocytes depend on the transporter ABCC4 to export cAMP as an extracellular signal. Extracellular cAMP, through its metabolite adenosine, reduced cardiomyocyte cAMP formation and hypertrophy by activating A1 adenosine receptors while delivering an antifibrotic signal to cardiac fibroblasts by A2 adenosine receptor activation. Together, our data reveal a paracrine role for secreted cAMP in intercellular signaling in the myocardium, and we postulate that secreted cAMP may also constitute an important signal in other tissues

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“…While the embryonic analyses have typically utilized 150-μm thick sections 2,4 , section thickness can be a great as 500 μm without evidence of oxygen deprivation 8 . These slice cultures have been maintained as long as two months in culture, with most slices maintaining contractility throughout this period 9 . Compared to studies in isolated cardiomyocytes, these slice cultures allow the co-culture of cardiomyocytes with their neighboring cell types and provide a useful method for ex vivo analysis.…”

Section: Introductionmentioning

confidence: 99%

A Novel <em>Ex vivo</em> Culture Method for the Embryonic Mouse Heart

Dyer

Patterson

2013

JoVE

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Citation: Dyer, L.A., Patterson, C. A Novel Ex vivo Culture Method for the Embryonic Mouse Heart. J. Vis. Exp. (75), e50359, doi:10.3791/50359 (2013). AbstractDevelopmental studies in the mouse are hampered by the inaccessibility of the embryo during gestation. Thus, protocols to isolate and culture individual organs of interest are essential to provide a method of both visualizing changes in development and allowing novel treatment strategies. To promote the long-term culture of the embryonic heart at late stages of gestation, we developed a protocol in which the excised heart is cultured in a semi-solid, dilute Matrigel. This substrate provides enough support to maintain the three-dimensional structure but is flexible enough to allow continued contraction. In brief, hearts are excised from the embryo and placed in a mixture of cold Matrigel diluted 1:1 with growth medium. After the diluted Matrigel solidifies, growth medium is added to the culture dish. Hearts excised as late as embryonic day 16.5 were viable for four days post-dissection. Analysis of the coronary plexus shows that this method does not disrupt coronary vascular development. Thus, we present a novel method for long-term culture of embryonic hearts. Video LinkThe video component of this article can be found at

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Organotypic slice culture from human adult ventricular myocardium

Abstract: Organotypic heart slices represent a multicellular model of the human myocardium and a novel platform for studies ranging from the investigation of molecular interactions to tissue engineering.

Cited by 93 publications

References 38 publications

Electrophysiological Characterization of Spontaneously Contracting Cell Aggregates Obtained from Rainbow Trout Larvae with Multielectrode Arrays

Electrophysiological Characterization of Spontaneously Contracting Cell Aggregates Obtained from Rainbow Trout Larvae with Multielectrode Arrays

Cardiac myocyte–secreted cAMP exerts paracrine action via adenosine receptor activation

A Novel <em>Ex vivo</em> Culture Method for the Embryonic Mouse Heart

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