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Updated: Jun 19, 2026
Isolation and Expansion of Human Glioblastoma Multiforme Tumor Cells Using the Neurosphere Assay
Published on: October 30, 2011
Isolation and partial characterization of a new human glioblastoma cell line
F M Brehar1, C Bleotu, L M Stefan
1Emergency Clinical Hospital Bagdasar-Arseni, Bucharest, Romania. felixbrehar@yahoo.com
Abstract:
Although significant progresses were made in the field of molecular biology of malignant cerebral gliomas, the prognostic of these tumors continues to be reserved. One of the therapeutic failure reasons is the incomplete knowledge regarding the origin of these tumors and cells features, which in fact represent an obstacle in developing a cell and molecular therapy guided against malignant cells responsible for the tumor development and for the therapeutic resistance. Initiation and characterization of glioblastoma cell lines represents an essential step in order to obtain a better in vitro and in vivo experimental model for glioblastoma. We describe here a new glioblastoma line, named T11, which was successfully isolated in our laboratories starting with a tumor sample obtained intraoperative from a 58 years-old female patient. The histopathological evaluation showed a grad IV WHO glioma (glioblastoma). The sample was prepared by manual fragmentation, followed by enzymatic digestions using different concentration of trypsin. The cell line has been cultivated for more than 150 passages. The characterization of the glioblastoma line consisted in the evaluation of cells proliferation capacity (growth curve), morphological features, karyotyping and identification of specific markers. We found that T11 expressed specific markers for glial progenitors and astrocytes (glial fibrillary acidic protein-GFAP); oligodendrocites (A2B5; O4), and microglia (CD45, CD 11b). Cells were negative for neuronal lineage markers like beta3-tubulin and NCAM. In order to evaluate the differentiation grade of T11 cell line, the presence of stem cell markers (nestin, CD133) was explored. T11l cells expressed higher level of nestin and lower level of CD133 comparing with standard glioblastoma cell line U87. T11 cell line expressed VEGF and Bcl-2, but not EGFR and Mdrl and Bax. This new line has distinct and unique characteristics when compared with standard glioblastoma cell line (e.g., U87) and may become a new and useful in vitro model for glioblastoma.
Insights
Researchers developed a new glioblastoma cell line, T11, from patient tumor samples. This T11 line exhibits unique characteristics, offering a valuable new model for studying glioblastoma and developing targeted therapies.
Area of Science:
- Neuro-oncology
- Cancer Cell Biology
- Molecular Oncology
Background:
- Malignant cerebral gliomas have a poor prognosis due to incomplete understanding of tumor origins and cell features.
- Developing effective cell and molecular therapies is hindered by limited knowledge of glioblastoma's malignant cells and therapeutic resistance mechanisms.
- Establishing well-characterized glioblastoma cell lines is crucial for advancing in vitro and in vivo experimental models.
Purpose of the Study:
- To isolate and characterize a novel glioblastoma cell line for use as an experimental model.
- To evaluate the potential of the new cell line in advancing glioblastoma research and therapeutic development.
Main Methods:
- Isolation of a new glioblastoma cell line (T11) from a patient tumor sample.
- Characterization including histopathology, cell proliferation assays, karyotyping, and specific marker analysis (GFAP, A2B5, O4, CD45, CD 11b, nestin, CD133, VEGF, Bcl-2, EGFR, Mdrl, Bax).
- Comparison of T11 cell line characteristics with a standard glioblastoma cell line (U87).
Main Results:
- The T11 cell line was successfully established and maintained for over 150 passages.
- T11 cells expressed markers for glial progenitors, astrocytes, oligodendrocytes, and microglia, but not neuronal markers.
- T11 cells showed higher nestin and lower CD133 expression compared to U87, and expressed VEGF and Bcl-2, but not EGFR, Mdrl, or Bax.
Conclusions:
- The newly isolated T11 glioblastoma cell line possesses distinct characteristics compared to established lines like U87.
- The T11 cell line represents a potentially valuable new in vitro model for glioblastoma research.
- Further investigation of T11 may aid in understanding glioblastoma biology and developing targeted therapies.
