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Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment
Published on: March 25, 2020
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Immunostaining protocol for infiltrating brain cancer spheroids for light-sheet imaging
Benedicte Bjørknes1, Oliver Emil Neye1, Petra Hamerlik2
1The Niels Bohr Institute, University of Copenhagen, Copenhagen, Denmark.
Plos One
|February 9, 2023
Summary
Researchers identified vimentin-rich cells on glioblastoma spheroid surfaces as key drivers of brain infiltration. This finding offers new insights into therapeutic resistance and guides future anti-cancer drug development targeting cytoskeleton reorganization.
Area of Science:
- Neuroscience
- Cell Biology
- Oncology
Background:
- Glioblastoma is an aggressive brain cancer characterized by rapid growth and infiltration.
- Therapeutic resistance and invasive growth contribute to the poor prognosis of glioblastoma patients.
- Understanding the mechanisms of glioblastoma cell infiltration is crucial for developing effective treatments.
Purpose of the Study:
- To develop a protocol for detecting vimentin in cells at the surface of proliferating glioblastoma spheroids.
- To investigate the role of cytoskeleton-reinforcing cells in initiating tumor infiltration.
- To provide a foundation for studying anti-cancer drug effects on cytoskeleton dynamics.
Main Methods:
- Combined a classical invasion assay with immunofluorescence.
- Utilized light-sheet imaging for high-resolution visualization.
- Focused on detecting the intermediate filament vimentin in spheroid surface cells.
Main Results:
- Identified vimentin-rich cells specifically at the proliferating spheroid surface.
- Demonstrated that these cytoskeleton-reinforcing cells initiate tumor infiltration.
- Established a correlation between vimentin expression and invasive behavior.
Conclusions:
- Vimentin-expressing cells on the spheroid surface are critical for initiating glioblastoma infiltration.
- The developed protocol enables detailed investigation of cytoskeleton dynamics in invasive cancer cells.
- Findings pave the way for targeted therapies aimed at inhibiting glioblastoma invasion and improving treatment outcomes.
