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Updated: May 9, 2026
Optimization of High Grade Glioma Cell Culture from Surgical Specimens for Use in Clinically Relevant Animal Models and 3D Immunochemistry
Published on: January 7, 2014
Heterogeneous reovirus susceptibility in human glioblastoma stem-like cell cultures
S K van den Hengel1, R K Balvers, I J C Dautzenberg
11] Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands [2] Department of Neuro-Oncology, Brain Tumor Center, Erasmus Medical Center, Rotterdam, The Netherlands.
Abstract:
Glioblastoma (GB) is a devastating disease for which new treatment modalities are needed. Efficacious therapy requires the removal of stem-cell like cells, these cells drive tumor progression because of their ability to self-renew and differentiate. In glioblastoma, the GB stem-like cells (GSC) form a small population of tumor cells and possess high resistance to chemo and radiation therapies. To assess the sensitivity of GSC to reovirus-mediated cytolysis, a panel of GSC cultures was exposed to wild-type reovirus Type 3 Dearing (T3D) and its junction adhesion molecule-A (JAM-A)-independent mutant, jin-1. Several parameters were evaluated, including the fraction of cells expressing the JAM-A reovirus receptor, the fraction of cells synthesizing reovirus proteins, the number of infectious reovirus particles required to reduce cell viability, the amount of infectious progeny reovirus produced and the capacity of the reoviruses to infect the GSC in 3-dimensional (3D) tumor cell spheroids. Our data demonstrate a marked heterogeneity in the susceptibility of the cultures to reovirus-induced cytolysis. While in monolayer cultures the jin-1 reovirus was generally more cytolytic than the wild-type reovirus T3D, in the 3D GSC spheroids, these viruses were equally effective. Despite the variation in reovirus sensitivity between the different GSC cultures, our data support the use of reovirus as an oncolytic agent. It remains to be established whether the variation in the reovirus sensitivity correlates with a patient's response to reovirus therapy. Moreover, our data show that the expression of the JAM-A receptor is not a major determinant of reovirus sensitivity in 3D GSC cultures.
Insights
Reovirus shows potential as an oncolytic agent against glioblastoma stem-like cells (GSCs). While sensitivity varies, reovirus effectively targets these resilient cells in 3D cultures, suggesting therapeutic promise.
Area of Science:
- Oncology
- Virology
- Cancer Stem Cell Biology
Background:
- Glioblastoma (GB) is an aggressive brain tumor with limited treatment options.
- Glioblastoma stem-like cells (GSCs) drive tumor progression and resist conventional therapies.
- Novel therapeutic strategies targeting GSCs are urgently needed.
Purpose of the Study:
- To evaluate the efficacy of reovirus-mediated cytolysis against glioblastoma stem-like cells (GSCs).
- To compare the activity of wild-type reovirus (T3D) and a junction adhesion molecule-A (JAM-A)-independent mutant (jin-1).
- To assess reovirus sensitivity in both 2D monolayer and 3D spheroid GSC cultures.
Main Methods:
- Exposure of GSC cultures to wild-type reovirus T3D and jin-1 mutant.
- Quantification of JAM-A receptor expression, viral protein synthesis, and infectious viral particles.
- Assessment of cell viability and progeny virus production in 2D and 3D GSC models.
Main Results:
- Significant heterogeneity observed in GSC susceptibility to reovirus-induced cell death.
- In 3D spheroids, both T3D and jin-1 reoviruses demonstrated comparable efficacy.
- JAM-A receptor expression was not a primary determinant of reovirus sensitivity in 3D GSC cultures.
Conclusions:
- Reovirus demonstrates oncolytic potential against glioblastoma stem-like cells.
- The study supports the use of reovirus as a potential therapeutic agent for glioblastoma.
- Further investigation is required to correlate reovirus sensitivity with patient treatment outcomes.