A normal adult heart is composed of several different cell types, among which cardiac mesenchymal stromal cells represent an abundant population. The isolation of these cells offers the possibility of studying their involvement in cardiac diseases, and, in addition, provides a useful primary cell model to investigate biological mechanisms. Here, the method for the isolation of C-MSC from arrhythmogenic cardiomyopathy patients’ bioptic samples is described. The endomyocardial biopsy sampling is guided in the right ventricular areas adjacent to the scar visualized by electro-anatomical mapping. The digestion of the biopsies in collagenase and their plating on a plastic dish in culture medium to allow C-MSC growth is described. The isolated cells can be expanded in culture for several passages. To confirm their mesenchymal phenotype, the description of immuno-phenotypical characterization is provided. C-MSC are able to differentiate into several cell types like adipocytes, chondrocytes, and osteoblasts: in the context of ACM, characterized by adipocyte deposits in patients’ hearts, the protocols for the adipogenic differentiation of C-MSC and the characterization of lipid droplet accumulation are described.

Isolation and characterization of cardiac mesenchymal stromal cells from endomyocardial bioptic samples of arrhythmogenic cardiomyopathy patients / C.A. Pilato, I. Stadiotti, A.S. Maione, V. Saverio, V. Catto, F. Tundo, A.D. Russo, C. Tondo, G. Pompilio, M. Casella, E. Sommariva. - In: JOURNAL OF VISUALIZED EXPERIMENTS. - ISSN 1940-087X. - 2018:132(2018 Feb). [10.3791/57263]

Isolation and characterization of cardiac mesenchymal stromal cells from endomyocardial bioptic samples of arrhythmogenic cardiomyopathy patients

C.A. Pilato
Primo
;
I. Stadiotti
Secondo
;
F. Tundo;A.D. Russo;C. Tondo;G. Pompilio
Penultimo
;
2018-02

Abstract

A normal adult heart is composed of several different cell types, among which cardiac mesenchymal stromal cells represent an abundant population. The isolation of these cells offers the possibility of studying their involvement in cardiac diseases, and, in addition, provides a useful primary cell model to investigate biological mechanisms. Here, the method for the isolation of C-MSC from arrhythmogenic cardiomyopathy patients’ bioptic samples is described. The endomyocardial biopsy sampling is guided in the right ventricular areas adjacent to the scar visualized by electro-anatomical mapping. The digestion of the biopsies in collagenase and their plating on a plastic dish in culture medium to allow C-MSC growth is described. The isolated cells can be expanded in culture for several passages. To confirm their mesenchymal phenotype, the description of immuno-phenotypical characterization is provided. C-MSC are able to differentiate into several cell types like adipocytes, chondrocytes, and osteoblasts: in the context of ACM, characterized by adipocyte deposits in patients’ hearts, the protocols for the adipogenic differentiation of C-MSC and the characterization of lipid droplet accumulation are described.
Adipogenesis; Arrhythmogenic Cardiomyopathy; ARVC; Cardiac mesenchymal stromal cells; Characterization; Developmental biology; Differentiation; Endomyocardial biopsy; Isolation; Issue 132; Adult; Biopsy; Cardiomyopathies; Cell Differentiation; Cells, Cultured; Humans; Mesenchymal Stromal Cells; Myocytes, Cardiac; Neuroscience (all); Chemical Engineering (all); Biochemistry, Genetics and Molecular Biology (all); Immunology and Microbiology (all)
Settore MED/11 - Malattie dell'Apparato Cardiovascolare
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/585788
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