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Isolation,Culture, and Functional Characterization of Adult Mouse Cardiomyoctyes
Authors:Evan Lee Graham  Cristina Balla  Hannabeth Franchino  Yonathan Melman  Federica del Monte  Saumya Das
Affiliation:1.Cardiovascular Institute, Beth Israel Deaconess Medical Center, Harvard Medical School;2.Division of Cardiology, Sapienza University
Abstract:
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.
Keywords:Cellular Biology   Issue 79   Medicine   Cardiology   Cellular Biology   Anatomy   Physiology   Mice   Ion Channels   Primary Cell Culture   Cardiac Electrophysiology   adult mouse cardiomyocytes   cell isolation   IonOptix   Cell Culture   adenoviral transfection   patch clamp   fluorescent nanosensor
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