Characterization of Immune Cells in Human Adipose Tissue by Using Flow Cytometry

Suzan Wetzels, Mitchell Bijnen, Erwin Wijnands, Erik A. L. Biessen, Casper G. Schalkwijk, Kristiaan Wouters*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

3 Citations (Web of Science)

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.
Original languageEnglish
Article numbere57319
Number of pages9
JournalJournal of visualized experiment
Issue number133
DOIs
Publication statusPublished - 1 Mar 2018

Keywords

  • Medicine
  • Issue 133
  • Flow cytometry
  • human
  • obesity
  • immune cells
  • macrophages
  • adipose tissue
  • INSULIN-RESISTANCE
  • MAST-CELLS
  • OBESITY
  • INFLAMMATION
  • MACROPHAGES
  • CONTRIBUTES
  • DISEASE

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