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A Simple and Efficient Method to Isolate Macrophages from Mixed Primary Cultures of Adult Liver Cells
Published on: May 24, 2011
Human Liver Macrophage Subsets Defined by CD32
Xia Wu1, Nicole Hollingshead1, Jessica Roberto1
1Departments of Pathology, University of Washington Medical Center, Seattle, WA, United States.
Abstract:
Human liver myeloid cells are imperfectly defined, but it is broadly agreed that cells of stellate appearance in situ, expressing the markers CD11b and CD68, are the liver's resident macrophages, classically termed Kupffer cells. Recent investigations using single cell RNA sequencing and unsupervised clustering algorithms suggest there are two populations of cells with the characteristics of tissue macrophages in human liver. We therefore analyzed dissociated human liver tissue using the markers CD11b and CD68 to define macrophage-like cells and found within this population two subsets that differ in their expression of multiple surface markers. These subsets were FACS-sorted based on CD32 expression, and gene expression analysis identified them with human liver myeloid cell subsets that were previously defined by two independent single cell RNA sequencing studies. Using qRT-PCR we found that the two subsets differed in the expression of genes associated with T cell activation and immunosuppression, suggesting distinct roles in T cell tolerance. In addition, one subset expressed two markers, CD1C and CD11c, more often seen on classical dendritic cells. Criteria used to distinguish macrophages from dendritic cells in other tissues may need to be revised in the human liver.
Insights
Researchers identified two distinct subsets of human liver myeloid cells, differing in surface markers and gene expression. These subsets suggest unique roles in T cell tolerance and may challenge current definitions of liver macrophages and dendritic cells.
Area of Science:
- Immunology
- Cell Biology
- Hepatology
Background:
- Human liver myeloid cells, including Kupffer cells (liver macrophages), are not fully characterized.
- Previous single-cell RNA sequencing studies suggest heterogeneity within liver macrophage populations.
Purpose of the Study:
- To define and characterize distinct subsets of macrophage-like cells in human liver.
- To investigate the potential functional differences between these subsets, particularly in T cell interactions.
Main Methods:
- Analysis of dissociated human liver tissue using CD11b and CD68 markers.
- Fluorescence-activated cell sorting (FACS) based on CD32 expression to isolate subsets.
- Gene expression analysis (qRT-PCR) to identify subset characteristics and functions.
Main Results:
- Two distinct subsets of CD11b+CD68+ myeloid cells were identified in the human liver.
- These subsets exhibited differential expression of surface markers and genes related to T cell activation and immunosuppression.
- One subset expressed CD1C and CD11c, markers typically associated with classical dendritic cells.
Conclusions:
- The human liver contains at least two distinct populations of myeloid cells with macrophage characteristics.
- These subsets may play different roles in regulating T cell responses and maintaining immune tolerance.
- Current criteria for distinguishing macrophages from dendritic cells in the human liver may require re-evaluation.
