Isolation and Functional Characterization of Human Ventricular Cardiomyocytes from Fresh Surgical Samples

Coppini, Raffaele; Ferrantini, Cecila; Aiazzi, Alessandro; Mazzoni, Luca; Sartiani, Laura; Mugelli, Alessandro; Poggesi, Corrado; Cerbai, Elisabetta

doi:10.3791/51116

Cited by 37 publications

(30 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Protocols for isolating viable myocytes from small surgical samples exist and involve enzymatic digestion of the tissue fragments excised during elective surgical procedures followed by purification, resuspension, and Ca 2+ readaptation of the cells. 241,242 These human myocytes are mostly used to characterize specific functional (electrophysiological and Ca 2+ handling) abnormalities in patients. The small yield of cells (because of the limited size of the biopsy specimen) limits their use for assessment of metabolic activity until a highly sensitive single-cell technique becomes available.…”

Section: Metabolism Of Isolated Cardiac Cells and Cell Linesmentioning

confidence: 99%

Assessing Cardiac Metabolism

Taegtmeyer¹,

Young²,

Lopaschuk³

et al. 2016

Circulation Research

237

147

View full text Add to dashboard Cite

In a complex system of interrelated reactions, the heart converts chemical energy to mechanical energy. Energy transfer is achieved through coordinated activation of enzymes, ion channels, and contractile elements, as well as structural and membrane proteins. The heart’s needs for energy are difficult to overestimate. At a time when the cardiovascular research community is discovering a plethora of new molecular methods to assess cardiac metabolism, the methods remain scattered in the literature. The present statement on “Assessing Cardiac Metabolism” seeks to provide a collective and curated resource on methods and models used to investigate established and emerging aspects of cardiac metabolism. Some of those methods are refinements of classic biochemical tools, whereas most others are recent additions from the powerful tools of molecular biology. The aim of this statement is to be useful to many and to do justice to a dynamic field of great complexity.

show abstract

Section: Metabolism Of Isolated Cardiac Cells and Cell Linesmentioning

confidence: 99%

Assessing Cardiac Metabolism

Taegtmeyer¹,

Young²,

Lopaschuk³

et al. 2016

Circulation Research

237

147

View full text Add to dashboard Cite

show abstract

“…Surgical septal specimens from HCM and control patients were washed with standard cardioplegic solution and processed within 30 min from excision. Endocardial trabeculae suitable for mechanical measurements were dissected, and the remaining tissue was minced and subjected to enzymic dissociation to obtain viable single myocytes, as previously described (Coppini et al, ).…”

Section: Methodsmentioning

confidence: 99%

Late sodium current inhibitors to treat exercise‐induced obstruction in hypertrophic cardiomyopathy: an in vitro study in human myocardium

Ferrantini

Pioner

Mazzoni

et al. 2018

British J Pharmacology

Self Cite

View full text Add to dashboard Cite

Background and PurposeIn 30–40% of hypertrophic cardiomyopathy (HCM) patients, symptomatic left ventricular (LV) outflow gradients develop only during exercise due to catecholamine‐induced LV hypercontractility (inducible obstruction). Negative inotropic pharmacological options are limited to β‐blockers or disopyramide, with low efficacy and tolerability. We assessed the potential of late sodium current (INaL)‐inhibitors to treat inducible obstruction in HCM.Experimental ApproachThe electrophysiological and mechanical responses to β‐adrenoceptor stimulation were studied in human myocardium from HCM and control patients. Effects of INaL‐inhibitors (ranolazine and GS‐967) in HCM samples were investigated under conditions simulating rest and exercise.Key ResultsIn cardiomyocytes and trabeculae from 18 surgical septal samples of patients with obstruction, the selective INaL‐inhibitor GS‐967 (0.5 μM) hastened twitch kinetics, decreased diastolic [Ca2+] and shortened action potentials, matching the effects of ranolazine (10μM). Mechanical responses to isoprenaline (inotropic and lusitropic) were comparable in HCM and control myocardium. However, isoprenaline prolonged action potentials in HCM myocardium, while it shortened them in controls. Unlike disopyramide, neither GS‐967 nor ranolazine reduced force at rest. However, in the presence of isoprenaline, they reduced Ca2+‐transient amplitude and twitch tension, while the acceleration of relaxation was maintained. INaL‐inhibitors were more effective than disopyramide in reducing contractility during exercise. Finally, INaL‐inhibitors abolished arrhythmias induced by isoprenaline.Conclusions and ImplicationsRanolazine and GS‐967 reduced septal myocardium tension during simulated exercise in vitro and therefore have the potential to ameliorate symptoms caused by inducible obstruction in HCM patients, with some advantages over disopyramide and β‐blockers.

show abstract

“…; Coppini et al . ). Interestingly, in cTnT Δ160E ventricular cardiomyocytes, we observed about 20% of AP‐failing T‐tubules and consequent local Ca 2+ release abnormalities (Fig.…”

Section: Physiology Of the Transverse‐axial Tubular Systemmentioning

confidence: 97%

Electrical defects of the transverse‐axial tubular system in cardiac diseases

Crocini

Ferrantini

Coppini

et al. 2017

The Journal of Physiology

Self Cite

View full text Add to dashboard Cite

Electrical excitability is an essential feature of cardiomyocytes and the homogenous propagation of the action potential is guaranteed by a complex network of membrane invaginations called the transverse-axial tubular system (TATS). TATS structural remodelling is Claudia Crocini did her graduate work at the European Laboratory for Non Linear Spectroscopy in Florence, Italy. She studied the consequences of T-tubular electrical failure on Ca 2+ release and contraction in isolated cardiomyocytes from different pathological models. a hallmark of cardiac diseases and we demonstrated that this can be accompanied by electrical defects at single T-tubular level. Using a random-access multi-photon (RAMP) microscope, we found that pathological T-tubules can fail to conduct action potentials, which delays local Ca 2+ release. Although the underlying causes for T-tubular electrical failure are still unknown, our findings suggest that they are likely to be related to local ultrastructural alterations. Here, we first review the experimental approach that allowed us to observe and dissect the consequences of TATS electrical dysfunction and then propose two different strategies to unveil the reasons for T-tubular electrical failures. The first strategy consists in a correlative approach, in which the failing T-tubule identified with the RAMP microscope is then imaged with electron microscopy. The second approach exploits the diffusion of molecules within TATS to gain insights into the local TATS structure, even without a thorough reconstruction of the tubular network. Although challenging, the local electrical failure occurring at single T-tubules is a fundamental question that needs to be addressed and could provide novel insights in cardiac pathophysiology. Abstract figure legend The electrical defect of T-tubules and its consequence on Ca 2+ release. Action potential failure and Ca 2+ release in T-tubules of heart failure.

show abstract

Isolation and Functional Characterization of Human Ventricular Cardiomyocytes from Fresh Surgical Samples

Cited by 37 publications

References 27 publications

Assessing Cardiac Metabolism

Assessing Cardiac Metabolism

Late sodium current inhibitors to treat exercise‐induced obstruction in hypertrophic cardiomyopathy: an in vitro study in human myocardium

Electrical defects of the transverse‐axial tubular system in cardiac diseases

Contact Info

Product

Resources

About