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Updated: Feb 21, 2026
A Brain Tumor/Organotypic Slice Co-culture System for Studying Tumor Microenvironment and Targeted Drug Therapies
Published on: November 7, 2015
Organotypic brain explant culture as a drug evaluation system for malignant brain tumors
Noriaki Minami1,2, Yusuke Maeda1, Shunsuke Shibao1
1Division of Gene Regulation, Institute for Advanced Medical Research, Keio University School of Medicine, Tokyo, Japan.
Abstract:
Therapeutic options for malignant brain tumors are limited, with new drugs being continuously evaluated. Organotypic brain slice culture has been adopted for neuroscience studies as a system that preserves brain architecture, cellular function, and the vascular network. However, the suitability of brain explants for anticancer drug evaluation has been unclear. We here adopted a mouse model of malignant glioma based on expression of H-RasV12 in Ink4a/Arf-/- neural stem/progenitor cells to establish tumor-bearing brain explants from adult mice. We treated the slices with cisplatin, temozolomide, paclitaxel, or tranilast and investigated the minimal assays required to assess drug effects. Serial fluorescence-based tumor imaging was sufficient for evaluation of cisplatin, a drug with a pronounced cytotoxic action, whereas immunostaining of cleaved caspase 3 (a marker of apoptosis) and of Ki67 (a marker of cell proliferation) was necessary for the assessment of temozolomide action and immunostaining for phosphorylated histone H3 (a marker of mitosis) allowed visualization of paclitaxel-specific effects. Staining for cleaved caspase 3 was also informative in the assessment of drug toxicity for normal brain tissue. Incubation of explants with fluorescently labeled antibodies to CD31 allowed real-time imaging of the microvascular network and complemented time-lapse imaging of tumor cell invasion into surrounding tissue. Our results suggest that a combination of fluorescence imaging and immunohistological staining allows a unified assessment of the effects of various classes of drug on the survival, proliferation, and invasion of glioma cells, and that organotypic brain slice culture is therefore a useful tool for evaluation of antiglioma drugs.
Insights
Organotypic brain slice culture effectively models malignant glioma in mice. This method, using fluorescence imaging and immunostaining, allows comprehensive evaluation of antiglioma drug efficacy and toxicity.
Area of Science:
- Neuroscience
- Oncology
- Drug Discovery
Background:
- Malignant brain tumors have limited therapeutic options.
- Organotypic brain slice culture preserves brain architecture and function but its use for anticancer drug evaluation is unclear.
Purpose of the Study:
- To establish tumor-bearing brain explants from a mouse glioma model.
- To investigate minimal assays for assessing antiglioma drug effects in brain explants.
Main Methods:
- Established H-RasV12-expressing glioma in Ink4a/Arf-/- neural stem/progenitor cells in adult mice.
- Prepared tumor-bearing brain explants and treated with cisplatin, temozolomide, paclitaxel, or tranilast.
- Utilized serial fluorescence imaging, cleaved caspase 3, Ki67, phosphorylated histone H3, and CD31 immunostaining.
Main Results:
- Cisplatin effects assessed by fluorescence imaging; temozolomide by apoptosis and proliferation markers.
- Paclitaxel effects visualized by mitosis markers; drug toxicity evaluated using cleaved caspase 3.
- Microvascular network and tumor invasion imaged in real-time, complementing drug effect assessments.
Conclusions:
- A combination of imaging and immunostaining enables unified assessment of antiglioma drug effects.
- Organotypic brain slice culture is a valuable tool for evaluating antiglioma drugs.
