Multiparametric Flow Cytometry-Based Immunophenotyping of Mouse Liver Immune Cells

0563508 - ÚOCHB 2023 RIV CH eng J - Journal Article
Vaneková, Lenka - Pimková Polidarová, Markéta - Veverka, Václav - Birkuš, Gabriel - Brázdová, Andrea
Multiparametric Flow Cytometry-Based Immunophenotyping of Mouse Liver Immune Cells.
Methods and Protocols. Roč. 5, č. 5 (2022), č. článku 70. E-ISSN 2409-9279
R&D Projects: GA MŠMT(CZ) EF16_019/0000729
Institutional support: RVO:61388963
Keywords : flow cytometry * immunophenotyping * mouse liver * PBS-based liver perfusion * non-parenchymal cells
OECD category: Biochemistry and molecular biology
Impact factor: 2.4, year: 2022
Method of publishing: Open access
https://doi.org/10.3390/mps5050070

The liver is a complex organ that governs many types of metabolisms, including energy metabolism and other cellular processes. The liver also plays a crucial role in important functions in immunity, and the activity of liver tissue-associated immunity affects the outcome of many liver pathologies. A thorough characterization of the liver immune microenvironment may contribute to a better understanding of immune signaling, the mechanisms of specific immune responses, and even to improved predictions about therapy outcomes. In this paper, we present an optimized, simple, and rapid protocol to characterize the liver-associated immune cell milieu. We believe that the most suitable technique for obtaining a complex immune cell suspension and for removing contaminating blood cells is to perform mouse liver perfusion, using only phosphate buffer saline. Combining an enzymatic digestion and a mechanical dissociation of liver tissue, followed by cell purification, improves downstream applications. This combination is an essential prerequisite for immune cell determination and characterization. We then demonstrate a flow cytometry-based multiparametric immunophenotyping along with a gating strategy to detect and quantify liver endothelial cells, T cells (helper and cytotoxic), B cells, NK cells, NKT cells, neutrophils, monocytes (subsets included), dendritic cells (subsets included), macrophages and Kupffer cells.
Permanent Link: https://hdl.handle.net/11104/0335459