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Title: | Characterization of Immune Cells in Human Adipose Tissue by Using Flow Cytometry | Authors: | WETZELS, Suzan Bijnen, Mitchell Wijnands, Erwin Biessen, Erik A. L. Schalkwijk, Casper G. WOUTERS, Kristiaan |
Issue Date: | 2018 | Publisher: | JOURNAL OF VISUALIZED EXPERIMENTS | Source: | JOVE-JOURNAL OF VISUALIZED EXPERIMENTS,(133) (Art N° e57319) | Abstract: | Infiltration of immune cells in the subcutaneous and visceral adipose tissue (AT) deposits leads to a low-grade inflammation contributing to the development of obesity-associated complications such as type 2 diabetes. To quantitatively and qualitatively investigate the immune cell subsets in human AT deposits, we have developed a flow cytometry approach. The stromal vascular fraction (SVF), containing the immune cells, is isolated from subcutaneous and visceral AT biopsies by collagenase digestion. Adipocytes are removed after centrifugation. The SVF cells are stained for multiple membrane-bound markers selected to differentiate between immune cell subsets and analyzed using flow cytometry. As a result of this approach, pro-and anti-inflammatory macrophage subsets, dendritic cells (DCs), B-cells, CD4(+) and CD8(+) T-cells, and NK cells can be detected and quantified. This method gives detailed information about immune cells in AT and the amount of each specific subset. Since there are numerous fluorescent antibodies available, our flow cytometry approach can be adjusted to measure various other cellular and intracellular markers of interest. | Notes: | [Wetzels, Suzan; Bijnen, Mitchell; Schalkwijk, Casper G.; Wouters, Kristiaan] MUMC, Dept Internal Med, Maastricht, Netherlands. [Wetzels, Suzan; Bijnen, Mitchell; Wijnands, Erwin; Biessen, Erik A. L.; Schalkwijk, Casper G.; Wouters, Kristiaan] MUMC, CARIM, Maastricht, Netherlands. [Wetzels, Suzan] Hasselt Univ, Biomed Res Inst, Dept Immunol & Biochem, Hasselt, Belgium. [Wijnands, Erwin; Biessen, Erik A. L.] MUMC, Dept Pathol, Maastricht, Netherlands. | Keywords: | Medicine; Issue 133; Flow cytometry; human; obesity; immune cells; macrophages; adipose tissue;Medicine; Issue 133; Flow cytometry; human; obesity; immune cells; macrophages; adipose tissue | Document URI: | http://hdl.handle.net/1942/27713 | ISSN: | 1940-087X | e-ISSN: | 1940-087X | DOI: | 10.3791/57319 | ISI #: | 000443329500103 | Rights: | Copyright © 2018 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2019 |
Appears in Collections: | Research publications |
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wetzels 1.pdf Restricted Access | Published version | 511.39 kB | Adobe PDF | View/Open Request a copy |
Characterization of immune cells in human AT.pdf | Peer-reviewed author version | 654.68 kB | Adobe PDF | View/Open |
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