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Updated: Jun 8, 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
Functional and molecular characterization of glioblastoma multiforme-derived cancer stem cells
C Tomuleasa1, O Soritau, D Rus-Ciuca
1Department of Cancer Immunology, Ion Chiricuta Comprehensive Cancer Center, Cluj Napoca, Romania. ciprian.tomuleasa@gmail.com
Purpose:
Brain tumors are the leading cause of cancer mortality in children and remain incurable despite advances in surgery and adjuvant therapies. The failure of malignant gliomas to respond to conventional treatment reflects the unique biology of these tumors, linked to a small population of stem-like precursors. This study describes the characteristics of stem cells isolated from glioblastoma multiforme (GM) and gives insight into the mechanism of brain tumorigenesis.
Methods:
Tumor stem-like precursors were identified from primary human GM-derived cell culture using immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). Cells were cultured in vitro in stem cell medium supplemented with growth factors and then the capacity of the surviving stem-like precursors to form tumor spheres and to continue to proliferate after chemoradiotherapy were tested.
Results:
The tumor cells expressed the cellular markers CD133, CD105, CD90, Nanog, Oct 3/4, CXCR4, nestin, glial fibrillary acidic protein (GFAP), neurofilament protein (NF) and human glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Cells also displayed a high proliferative potential despite chemotherapy and irradiation and also had the ability to form spheroids in suspension.
Conclusion:
High grade gliomas contain stem-like precursors, which exhibit neural stem cell properties with tumorigenicity, establishing a novel developmental paradigm in the study of brain carcinogenesis and providing a powerful tool to develop patient-tailored therapy for this devastating disease.
Insights
Pediatric brain tumors, particularly glioblastoma multiforme (GM), contain stem-like cells. These cells resist treatment and drive tumor growth, offering new therapeutic targets.
Area of Science:
- Oncology
- Neuroscience
- Stem Cell Biology
Background:
- Brain tumors are the leading cause of cancer mortality in children.
- Malignant gliomas are largely incurable due to their unique biology.
- A small population of stem-like precursors is implicated in treatment failure.
Purpose of the Study:
- To characterize stem cells isolated from glioblastoma multiforme (GM).
- To investigate the mechanism of brain tumorigenesis driven by these cells.
Main Methods:
- Identified tumor stem-like precursors from human GM cell cultures.
- Utilized immunocytochemistry and RT-PCR for cell identification.
- Assessed spheroid formation and proliferation capacity post-chemotherapy and irradiation.
Main Results:
- Isolated cells expressed key stem cell markers (CD133, Nanog, Oct 3/4) and neural markers (nestin, GFAP).
- These cells demonstrated high proliferative potential even after chemoradiotherapy.
- The cells exhibited the ability to form tumor spheres in vitro.
Conclusions:
- High-grade gliomas harbor stem-like precursors with neural stem cell properties and tumorigenicity.
- This finding establishes a new developmental model for brain cancer.
- These precursors represent a promising target for developing personalized therapies for glioblastoma.
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