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Updated: Aug 10, 2026
Isolation and Flow Cytometric Analysis of Glioma-infiltrating Peripheral Blood Mononuclear Cells
Published on: November 28, 2015
In vitro modulation of microglia motility by glioma cells is mediated by hepatocyte growth factor/scatter factor
B Badie1, J Schartner, J Klaver
1Department of Neurological Surgery, University of Wisconsin School of Medicine, Madison, USA.
Objective:
Considered as immune effector cells of the central nervous system, microglia represent a major component of the inflammatory cells found in malignant gliomas. Although their role in brain tumor biology is unclear, accumulation of microglia in malignant brain tumors may be mediated through active secretion of cytokines by glioma cells. Because hepatocyte growth factor/scatter factor (HGF/SF) has been shown to modulate glioma motility through an autocrine mechanism, and because microglia have been reported to express the HGF/SF receptor Met, we hypothesized that microglia recruitment by gliomas may also occur through the secretion of HGF/SF.
Methods:
The effect of glioma cells in augmenting BV-2 murine microglia motility was studied by using an in vitro Boyden chamber migration assay. To determine the chemokines involved in microglia migration, neutralizing monoclonal antibodies against monocyte chemotactic protein-1 and HGF/SF were tested. Immunoblotting was used to check for the expression of HGF/SF by glioma cells, and the expression of Met by BV-2 cells was examined by flow cytometry.
Results:
BV-2 migration was noted within 7 hours of incubation with both human (U251 MG and U373 MG) and murine (GL261) glioma cell lines. This migration corresponded to HGF/SF secretion by glioma cells and was completely inhibited by neutralizing monoclonal antibody against HGF/SF, but not monocyte chemotactic protein-1. Exposure of BV-2 cells to recombinant HGF/SF, but not monocyte chemotactic protein-1, resulted in their migration and down-regulation of Met in a dose-dependent fashion.
Conclusion:
HGF/SF, which plays a role in glioma motility and mitogenesis, may also act as a chemokine for microglia and may be responsible for the microglia infiltration in malignant gliomas. This active recruitment of microglia may play an important role in glioma biology.
Insights
Hepatocyte growth factor/scatter factor (HGF/SF) secreted by glioma cells attracts microglia, contributing to malignant glioma development. This finding highlights HGF/SF as a potential therapeutic target for brain tumors.
Area of Science:
- Neuroscience
- Immunology
- Oncology
Background:
- Microglia are immune cells in the central nervous system and are found in malignant gliomas.
- The precise role of microglia in brain tumor biology is not fully understood.
- Glioma cells may actively recruit microglia through cytokine secretion.
Purpose of the Study:
- To investigate the hypothesis that hepatocyte growth factor/scatter factor (HGF/SF) secreted by glioma cells recruits microglia.
- To explore the role of HGF/SF in microglia infiltration in malignant gliomas.
Main Methods:
- Utilized an in vitro Boyden chamber migration assay to assess microglia motility.
- Employed neutralizing monoclonal antibodies against monocyte chemotactic protein-1 and HGF/SF.
- Confirmed HGF/SF expression in glioma cells via immunoblotting and Met receptor expression in microglia using flow cytometry.
Main Results:
- Glioma cells (human and murine) induced significant BV-2 murine microglia migration.
- This migration was dependent on HGF/SF secretion by glioma cells and was inhibited by anti-HGF/SF antibodies.
- Recombinant HGF/SF induced microglia migration and Met receptor down-regulation.
Conclusions:
- HGF/SF acts as a chemokine, mediating microglia recruitment in malignant gliomas.
- Microglia infiltration, driven by HGF/SF, plays a significant role in glioma biology.
- HGF/SF may be a potential therapeutic target for managing malignant gliomas.
