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Multiparametric Flow Cytometry-Based Immunophenotyping of Mouse Liver Immune Cells
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SYSNO ASEP 0563508 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Multiparametric Flow Cytometry-Based Immunophenotyping of Mouse Liver Immune Cells Author(s) Vaneková, Lenka (UOCHB-X) ORCID
Pimková Polidarová, Markéta (UOCHB-X) ORCID
Veverka, Václav (UOCHB-X) RID, ORCID
Birkuš, Gabriel (UOCHB-X) ORCID
Brázdová, Andrea (UOCHB-X) ORCIDArticle number 70 Source Title Methods and Protocols. - : MDPI
Roč. 5, č. 5 (2022)Number of pages 15 s. Language eng - English Country CH - Switzerland Keywords flow cytometry ; immunophenotyping ; mouse liver ; PBS-based liver perfusion ; non-parenchymal cells OECD category Biochemistry and molecular biology R&D Projects EF16_019/0000729 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support UOCHB-X - RVO:61388963 UT WOS 000873086400001 EID SCOPUS 85140616880 DOI 10.3390/mps5050070 Annotation 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. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Year of Publishing 2023 Electronic address https://doi.org/10.3390/mps5050070
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