title: Isolation and functional characterization of human ventricular cardiomyocytes from fresh surgical samples.
abstract: Cardiomyocytes from diseased hearts are subjected to complex remodeling processes involving changes in cell structure, excitation contraction coupling and membrane ion currents. Those changes are likely to be responsible for the increased arrhythmogenic risk and the contractile alterations leading to systolic and diastolic dysfunction in cardiac patients. However, most information on the alterations of myocyte function in cardiac diseases has come from animal models. Here we describe and validate a protocol to isolate viable myocytes from small surgical samples of ventricular myocardium from patients undergoing cardiac surgery operations. The protocol is described in detail. Electrophysiological and intracellular calcium measurements are reported to demonstrate the feasibility of a number of single cell measurements in human ventricular cardiomyocytes obtained with this method. The protocol reported here can be useful for future investigations of the cellular and molecular basis of functional alterations of the human heart in the presence of different cardiac diseases. Further, this method can be used to identify novel therapeutic targets at cellular level and to test the effectiveness of new compounds on human cardiomyocytes, with direct translational value.
authors.affiliation: Department NeuroFarBa, Division of Pharmacology, University of Florence; raffaele.coppini@unifi.it.
authors.forename: Raffaele
authors.initials: R
authors.lastname: Coppini
authors.affiliation: Department of Clinical and Experimental Medicine, Division of Physiology, University of Florence.
authors.forename: Cecila
authors.initials: C
authors.lastname: Ferrantini
authors.affiliation: Department of Clinical and Experimental Medicine, Division of Physiology, University of Florence.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Aiazzi
authors.affiliation: Department NeuroFarBa, Division of Pharmacology, University of Florence.
authors.forename: Luca
authors.initials: L
authors.lastname: Mazzoni
authors.affiliation: Department NeuroFarBa, Division of Pharmacology, University of Florence.
authors.forename: Laura
authors.initials: L
authors.lastname: Sartiani
authors.affiliation: Department NeuroFarBa, Division of Pharmacology, University of Florence.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Mugelli
authors.affiliation: Department of Clinical and Experimental Medicine, Division of Physiology, University of Florence.
authors.forename: Corrado
authors.initials: C
authors.lastname: Poggesi
authors.affiliation: Department NeuroFarBa, Division of Pharmacology, University of Florence.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Cerbai
mesh_terms: D002118:Calcium / Q000032:analysis / Q000378:metabolism; D002312:Cardiomyopathy, Hypertrophic / Q000473:pathology / Q000601:surgery; D003584:Cytological Techniques / Q000379:methods*; D006352:Heart Ventricles / Q000166:cytology; D006801:Humans; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism / Q000502:physiology*; D018408:Patch-Clamp Techniques
chemical_list: D002118:Calcium