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Multiparametric Flow Cytometry-Based Immunophenotyping of Mouse Liver Immune Cells
Lenka Vanekova1,2, Marketa Pimkova Polidarova1, Vaclav Veverka1,2
1Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo namesti 542/2, 160 00 Prague, Czech Republic.
Methods and Protocols
|September 22, 2022
Summary
This study presents a rapid protocol for characterizing liver immune cells. The optimized method uses liver perfusion and enzymatic digestion for accurate immune cell analysis in mice.
Area of Science:
- Immunology
- Hepatology
- Cell Biology
Background:
- The liver's immune microenvironment is critical for its functions and disease outcomes.
- Understanding liver immunity aids in predicting therapeutic responses.
- Characterizing liver-associated immune cells is essential for research.
Purpose of the Study:
- To develop an optimized, simple, and rapid protocol for characterizing the liver immune cell milieu.
- To improve the accuracy of immune cell determination and characterization in liver tissue.
Main Methods:
- Mouse liver perfusion using phosphate buffer saline to obtain immune cell suspension.
- Enzymatic digestion and mechanical dissociation of liver tissue.
- Cell purification followed by multiparametric flow cytometry immunophenotyping.
Main Results:
- Demonstration of a protocol for detecting and quantifying various liver immune cells.
- Identification of liver endothelial cells, T cells, B cells, NK cells, NKT cells, neutrophils, monocytes, dendritic cells, macrophages, and Kupffer cells.
- Successful characterization of the liver immune microenvironment.
Conclusions:
- The presented protocol is effective for characterizing the liver immune microenvironment.
- This method facilitates a better understanding of immune signaling and responses in the liver.
- The protocol is a prerequisite for accurate immune cell determination and can aid in predicting therapy outcomes.
