Atrial Cardiomyocyte Isolation
Atrial Cardiomyocyte Isolation - Related Videos
11:33
Isolation and Kv Channel Recordings in Murine Atrial and Ventricular Cardiomyocytes
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Cited by 15 •
2013
Kv channel dysfunction is associated with cardiac arrhythmias. In order to study the molecular mechanisms that lead to such arrhythmias we utilize a systematic protocol for isolation of atrial and ventricular cardiomyocytes from Kv channel ancillary subunit knockout mice. Isolated cardiomyocytes can then immediately be used for cellular electrophysiological studies, biochemical or immunofluorescence (IF) assays.
08:52
Differentiation of Atrial Cardiomyocytes from Pluripotent Stem Cells Using the BMP Antagonist Grem2
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Cited by 6 •
2016
Generating cardiomyocytes from pluripotent stem cells in vitro allows access to large amounts of cardiac tissue in vitro for basic science and clinical applications. This protocol uses the atrializing factor Grem2 to both increase the numbers of cardiomyocytes obtained and to generate cardiomyocytes with an atrial phenotype.
07:50
Isolation and Culture of Neonatal Mouse Cardiomyocytes
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Cited by 187 •
2013
Primary mouse cardiomyocyte cultures are one of the pivotal tools for the investigation of myofibrillar organization and function. The following protocol describes the isolation and culture of primary cardiomyocytes from neonatal mouse hearts. The resulting cardiomyocyte cultures may be subsequently used for a variety of biomechanical, biochemical and cell-biological assays.
07:05
Isolation and Cryopreservation of Neonatal Rat Cardiomyocytes
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Cited by 49 •
2015
The isolation of neonatal rat cardiomyocytes is a time consuming and unpredictable procedure. This study describes methods for cryopreservation and thawing of neonatal rat cardiomyocytes that allows for more efficient use of cells. The thawed NRCMs can be used for various experiments without the need for performing isolations each time.
10:12
Isolation of Cardiomyocyte Nuclei from Post-mortem Tissue
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Cited by 61 •
2012
Cardiac nuclei are isolated via density sedimentation and immunolabeled with antibodies against pericentriolar material 1 (PCM-1) to identify and sort cardiomyocyte nuclei by flow cytometry.
FAQs
Gradual calcium restoration helps the preparation transition from digestion conditions toward conditions that support normal cardiomyocyte function. Because the isolation process disrupts tissue structure and uses calcium-sensitive cells, restoring calcium stepwise is part of preserving viability and contractile behavior. This makes subsequent measurements of calcium handling, contraction, and electrophysiology more interpretable.
Collagenase and protease address the extracellular matrix that holds atrial tissue together, while gentle mechanical dissociation helps release individual cells after enzymatic treatment. The balance matters: digestion must separate the tissue without undermining cell viability and contractile properties. Subsequent filtration removes larger remnants, producing a preparation more suitable for controlled cellular measurements.
These cells permit direct study of action potentials, ion currents, calcium handling, and contraction under controlled conditions. Investigators can also examine responses to drugs or disease-related stimuli at the cellular level. Because the preparation retains atrial cardiomyocyte properties, it connects measured electrical and mechanical behavior with questions about atrial electrophysiology and remodeling.
A typical workflow begins with enzymatic digestion of atrial tissue using collagenase and often protease. Gentle mechanical dissociation then separates cells, followed by filtration to remove tissue remnants. Calcium is restored gradually rather than all at once, and the resulting preparation is assessed through its viability and preserved contractile behavior before functional experiments.
Mechanical force is useful only after the extracellular matrix has been sufficiently disrupted. Gentle dissociation limits additional stress while releasing individual cardiomyocytes, helping maintain viability and contractile properties. This step directly affects whether the isolated cells remain suitable for measurements of electrical activity, calcium responses, contraction, and pharmacological effects.
Researchers can use the preparation to investigate atrial electrophysiology and arrhythmias, examine cellular changes associated with cardiac remodeling, and test responses to drugs or other disease-related stimuli. Its value lies in providing a native cellular system in which electrical, calcium-dependent, and contractile outcomes can be studied together during therapeutic development.