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Isolation and Physiological Analysis of Mouse Cardiomyocytes
Published on: September 7, 2014
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Isolation and physiological analysis of mouse cardiomyocytes
Gretchen M Roth1, David M Bader2, Elise R Pfaltzgraff3
1Department of Medicine, Vanderbilt University.
Journal of Visualized Experiments : Jove
|September 17, 2014
Summary
This study details a versatile method for isolating cardiomyocytes using Langendorff perfusion. This technique allows for detailed analysis of cellular function and is crucial for understanding heart failure at a cellular level.
Area of Science:
- Cardiology
- Cell Biology
- Physiology
Background:
- Cardiomyocytes are essential for heart function, generating contractile force.
- Studying isolated cardiomyocytes offers insights into heart physiology and disease.
- Existing cell culture methods lack critical in-vivo structures like organized sarcomeres.
Purpose of the Study:
- To present a refined protocol for isolating and analyzing cardiomyocytes.
- To investigate cellular changes associated with heart dysfunction, specifically dilated cardiomyopathy.
- To enable direct comparison of cardiomyocyte function under various experimental conditions.
Main Methods:
- Cardiomyocytes isolated from mouse hearts using Langendorff perfusion and enzymatic digestion.
- Cells are introduced to increasing calcium concentrations.
- Contractility analyzed using edge and sarcomere detection software; calcium transients visualized with fluorescent dyes.
Main Results:
- The protocol enables the study of live cardiomyocytes with known genetic backgrounds and heart function.
- Allows for reliable comparison of experimental conditions like genetic manipulation, drug treatment, and infection.
- Facilitates analysis of cytoskeletal elements through fixation and staining.
Conclusions:
- Langendorff perfusion is a versatile method for isolating cardiomyocytes.
- This technique enhances understanding of cellular mechanisms in heart failure.
- The protocol is valuable for comparing diverse experimental conditions and genetic models of cardiac disease.
