Isolation, Culture, and Live-Cell Imaging of Primary Rat Cardiomyocytes
Marina Leone1, Felix B Engel2,3
1Experimental Renal and Cardiovascular Research, Department of Nephropathology, Institute of Pathology, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Erlangen, Germany.
Insights
This guide details isolating and culturing rat cardiomyocytes for research. It provides methods for live-cell imaging, aiding cardiac regeneration and drug studies.
Area of Science:
- Cardiology
- Cell Biology
- Regenerative Medicine
Background:
- The heart comprises diverse cell types, including cardiomyocytes and non-myocytes, complicating in vivo studies.
- Interpreting in vivo data is challenging due to the heart's cellular complexity.
- The role of cardiomyocyte proliferation in cardiac regeneration remains unclear.
Purpose of the Study:
- To provide a practical guide for isolating and culturing primary neonatal and adult rat cardiomyocytes.
- To offer a detailed protocol for live-cell imaging of embryonic and neonatal cardiomyocytes.
- To highlight the importance of cell culture in cardiac research.
Main Methods:
- Isolation of primary neonatal and adult rat cardiomyocytes.
- Culture of isolated cardiomyocytes.
- Live-cell imaging techniques for cardiomyocytes.
Main Results:
- Established protocols for cardiomyocyte isolation and culture.
- Demonstrated feasibility of live-cell imaging for cardiomyocyte studies.
- Provided a resource for researchers in cardiac regeneration and pharmacology.
Conclusions:
- Cellular reductionist approaches using isolated cardiomyocytes are crucial for understanding cardiac function and disease.
- Cultured cardiomyocytes facilitate manipulation and analysis for research purposes.
- This guide supports advancements in cardiac regeneration, drug testing, and toxicological studies.
Abstract:
The heart is a complex organ consisting of a variety of different cardiomyocytes (ventricular vs. atrial, left vs. right ventricular, working vs. nodal) as well as other cell types, including endothelial cells and vascular smooth muscle cells. Pericytes, neurons, and immune cells are less abundant, yet still important. Whereas cardiomyocytes account for around 75% of the heart volume, 50-70% of the cells in the heart are non-myocytes. This complexity of the heart underlines the difficulties in interpreting data obtained in vivo. In the field of cardiac regeneration, it remains unclear whether it is possible to induce a significant number of cardiomyocytes to proliferate and whether the often-observed improvement in cardiac function after experimental therapies is due to the induction of cardiomyocyte proliferation. Therefore, the reductionist approach inherent to cultures of isolated cells continues to be of great importance, even though it is important to study heart disease in vivo due to interactions of the different cell types. Cultured cardiomyocytes allow for easy manipulation of cell behavior (e.g., cell division) and its analysis (e.g., live-cell imaging). In addition, isolated cells in culture are a valuable tool for pharmacological and toxicological studies. This chapter offers a practical guide to isolate and culture primary neonatal and adult rat cardiomyocytes and a detailed protocol for live-cell imaging of embryonic and neonatal cardiomyocytes.
