Cardiomyocyte Dissociation
Cardiomyocyte Dissociation - Related Videos
07:50
Isolation and Culture of Neonatal Mouse Cardiomyocytes
0 Views •
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.
08:42
Isolation, Culture and Transduction of Adult Mouse Cardiomyocytes
0 Views •
Cited by 35 •
2016
This protocol describes a step-by-step method for the reproducible isolation and long-term culture of adult mouse cardiomyocytes with high yield, purity, and viability.
07:05
Isolation and Cryopreservation of Neonatal Rat Cardiomyocytes
0 Views •
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.
09:41
Visualization of Cell Cycle Variations and Determination of Nucleation in Postnatal Cardiomyocytes
0 Views •
Cited by 5 •
2017
To distinguish cell division from cell cycle variations in cardiomyocytes, we present protocols using two transgenic mouse lines: Myh6-H2B-mCh transgenic mice, for the unequivocal identification of cardiomyocyte nuclei, and CAG-eGFP-anillin mice, for distinguishing cell division from cell cycle variations.
08:03
Simultaneous Assessment of Cardiomyocyte DNA Synthesis and Ploidy: A Method to Assist Quantification of Cardiomyocyte Regeneration and Turnover
0 Views •
Cited by 25 •
2016
Quantification of cardiomyocyte turnover is challenging. The protocol described here makes an important contribution to this challenge by enabling accurate and sensitive quantification of neo-cardiomyocyte nuclei generation and nuclei ploidy.
FAQs
The enzymatic step targets the structural connections that retain cardiac cells within heart tissue. Collagenase and related proteases break down extracellular matrix proteins and cell-cell attachments, reducing tissue cohesion around the cardiomyocytes. This biochemical loosening must be paired with physical processing, because digestion alone does not place the released cells into a usable suspension.
Gentle trituration mechanically separates cells after matrix digestion. Its purpose is to release cardiomyocytes without unnecessarily compromising viability or function. The resulting suspension supports measurements of contraction, calcium handling, electrical activity, metabolism, and responses to drugs or injury. Mechanical force therefore influences whether downstream observations reflect healthy isolated cells.
Isolated cells provide a direct experimental system for examining cell-level behavior outside the intact organ. Researchers can assess contraction, calcium handling, electrical activity, and metabolism, then evaluate responses to drugs or injury. This approach helps connect specific cardiomyocyte properties with broader questions in cardiovascular biology without requiring observations to remain within whole heart tissue.
A typical workflow begins with mincing heart tissue, followed by enzymatic digestion using collagenase and related proteases. Once extracellular matrix proteins and cell-cell attachments have been loosened, gentle trituration releases individual cardiomyocytes into suspension. The sequence matters because physical disruption is applied after biochemical weakening, helping obtain cells suitable for subsequent study.
Successful dissociation is not judged solely by obtaining a cell suspension. The isolated cardiomyocytes must remain viable and retain relevant functions, including contraction, calcium handling, and electrical activity. Preserving these properties determines whether the preparation can support meaningful experiments on metabolism, pharmacological responses, injury, or other cardiac processes.
Researchers use the resulting cells for primary cell culture, disease modeling, and studies of cardiac development and regeneration. The preparation also enables controlled investigation of drug responses, injury-related changes, contraction, calcium handling, electrical activity, and metabolism. These applications make dissociation a practical link between tissue processing and experiments focused on cardiac cell behavior.