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Updated: Dec 31, 2025
Isolation of Macrophage Subsets and Stromal Cells from Human and Mouse Myocardial Specimens
Published on: December 17, 2019
Isolation of Macrophage Subsets and Stromal Cells from Human and Mouse Myocardial Specimens
Geetika Bajpai1, Kory J Lavine2
1Department of Medicine, Washington University School of Medicine.
Insights
This study presents a simple protocol for extracting cardiac macrophages from mouse and human heart tissue. This method allows for detailed analysis of macrophage populations in various cardiac conditions.
Area of Science:
- Immunology
- Cardiovascular Biology
- Cellular Biology
Background:
- Macrophages are key immune cells in the heart, influencing inflammation and repair after injury.
- Understanding the diverse roles of cardiac macrophage subsets is crucial for cardiac research.
Purpose of the Study:
- To present a straightforward and reproducible protocol for isolating macrophages from cardiac tissue.
- To enable functional assays and molecular analyses (RNA sequencing) of cardiac macrophages.
Main Methods:
- Enzymatic digestion of myocardial tissue to create a single-cell suspension.
- Antibody staining followed by flow cytometry for cell identification and sorting.
- Application to both mouse and human cardiac specimens from healthy and diseased individuals.
Main Results:
- The protocol yields a single-cell suspension suitable for downstream analyses.
- The method is reliable, with minimal day-to-day variation.
- The technique is broadly applicable to various mouse models and human cardiac diseases.
Conclusions:
- This protocol provides a robust method for investigating cardiac macrophage heterogeneity.
- It facilitates research into the roles of different macrophage subsets in cardiac health and disease.
- The simplicity and wide applicability make it a valuable tool for the scientific community.
Abstract:
Macrophages represent the most heterogeneous and abundant immune cell populations in the heart and are central in driving inflammation and reparative responses after cardiac injury. How various subsets of macrophages orchestrate the immune responses after cardiac injury is an active area of research. Presented here is a simple protocol that our lab performs routinely, for the extraction of macrophages from mouse and human myocardium specimens obtained from healthy and diseased individuals. Briefly, this protocol involves enzymatic digestion of cardiac tissue to generate a single cell suspension, followed by antibody staining, and flow cytometry. This technique is suitable for functional assays performed on sorted cells as well as bulk and single cell RNA sequencing. A major advantage of this protocol is its simplicity, minimal day to day variation and wide applicability allowing investigation of macrophage heterogeneity across various mouse models and human disease entities.
