Isolation of murine hepatic myeloid cells with high yield and purity using immunomagnetic beads for subset analysis
Yung-Tsung Li1, Hui-Lin Wu2, Li-Ling Wu3
1Graduate Institute of Clinical Medicine, National Taiwan University College of Medicine, Taipei, Taiwan.
Insights
This study introduces an efficient method for isolating hepatic myeloid cells, including Kupffer cells and Ly6C+ monocytes/macrophages, preserving their function and phenotype. The new protocol yields pure, viable cells suitable for research.
Area of Science:
- Immunology
- Cell Biology
- Hepatology
Background:
- Isolating hepatic myeloid cells is crucial for understanding liver immunity.
- Existing methods often compromise cell phenotype and function.
- Kupffer cells and Ly6C+ monocytes/macrophages are key hepatic myeloid populations.
Purpose of the Study:
- To develop a straightforward and efficient method for isolating hepatic myeloid cells.
- To preserve the phenotypic and functional characteristics of isolated cells.
- To obtain large quantities of purified, viable, and functional hepatic myeloid cells.
Main Methods:
- Liver perfusion with collagenase.
- Purification using immunomagnetic particles.
- Isolation of Kupffer cells and Ly6C+ monocytes/macrophages.
Main Results:
- Successful isolation of hepatic myeloid cells, including Kupffer cells and Ly6C+ monocytes/macrophages.
- High yield of purified and viable cells.
- Preservation of cellular phenotype and function.
Conclusions:
- The described protocol provides an efficient means to isolate functional hepatic myeloid cells.
- This method is suitable for various research applications requiring high-quality immune cells from the liver.
- The protocol ensures cells remain physiologically relevant for downstream analyses.
Abstract:
There are numerous established techniques for isolating hepatic myeloid cells; however, preserving their phenotypic and functional characteristics can be challenging. We present a straightforward and efficient method to isolate hepatic myeloid cells, including Kupffer cells and lymphocyte antigen 6 complex, locus C+ (Ly6C+) monocytes/macrophages. The procedure involves perfusion of the liver with collagenase and purification with immunomagnetic particles. This protocol ensures the isolation of large quantities of purified, viable, and functional cells without influencing their physiological characteristics. For complete details on the use and execution of this protocol, please refer to Wu et al. (2019).1.
