Isolation, culture, and functional characterization of adult mouse cardiomyoctyes

The use of primary cardiomyocytes (CMs) in culture has provided a powerful complement to murine models of heart disease in advancing our understanding of heart disease. In particular, the ability to study ion homeostasis, ion channel function, cellular excitability and excitation-contraction coupling and their alterations in diseased conditions and by disease-causing mutations have led to significant insights into cardiac diseases. Furthermore, the lack of an adequate immortalized cell line to mimic adult CMs, and the limitations of neonatal CMs (which lack many of the structural and functional biomechanics characteristic of adult CMs) in culture have hampered our understanding of the complex interplay between signaling pathways, ion channels and contractile properties in the adult heart strengthening the importance of studying adult isolated cardiomyocytes. Here, we present methods for the isolation, culture, manipulation of gene expression by adenoviral-expressed proteins, and subsequent functional analysis of cardiomyocytes from the adult mouse. The use of these techniques will help to develop mechanistic insight into signaling pathways that regulate cellular excitability, Ca2+ dynamics and contractility and provide a much more physiologically relevant characterization of cardiovascular disease.

Data di pubblicazione: 2013
Titolo: Isolation, culture, and functional characterization of adult mouse cardiomyoctyes
Autori: Graham, Evan Lee; Balla, Cristina; Franchino, Hannabeth; Melman, Yonathan; del Monte, Federica; Das, Saumya
Rivista: JOURNAL OF VISUALIZED EXPERIMENTS
Parole Chiave: Adenoviral transfection; Adult mouse cardiomyocytes; Anatomy; Cardiac electrophysiology; Cardiology; Cell culture; Cell isolation; Cellular biology; Fluorescent nanosensor; Ion channels; Ionoptix; Issue 79; Medicine; Mice; Patch clamp; Physiology; Primary cell culture; Animals; Cytological Techniques; Mice; Myocytes, Cardiac; Patch-Clamp Techniques; Transfection; Biochemistry, Genetics and Molecular Biology (all); Chemical Engineering (all); Immunology and Microbiology (all); Neuroscience (all); Medicine (all)
Abstract in inglese: The use of primary cardiomyocytes (CMs) in culture has provided a powerful complement to murine models of heart disease in advancing our understanding of heart disease. In particular, the ability to study ion homeostasis, ion channel function, cellular excitability and excitation-contraction coupling and their alterations in diseased conditions and by disease-causing mutations have led to significant insights into cardiac diseases. Furthermore, the lack of an adequate immortalized cell line to mimic adult CMs, and the limitations of neonatal CMs (which lack many of the structural and functional biomechanics characteristic of adult CMs) in culture have hampered our understanding of the complex interplay between signaling pathways, ion channels and contractile properties in the adult heart strengthening the importance of studying adult isolated cardiomyocytes. Here, we present methods for the isolation, culture, manipulation of gene expression by adenoviral-expressed proteins, and subsequent functional analysis of cardiomyocytes from the adult mouse. The use of these techniques will help to develop mechanistic insight into signaling pathways that regulate cellular excitability, Ca2+ dynamics and contractility and provide a much more physiologically relevant characterization of cardiovascular disease.
Digital Object Identifier (DOI): 10.3791/50289
Handle: http://hdl.handle.net/11392/2376338
Appare nelle tipologie:03.1 Articolo su rivista

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