A simplified and modified procedure to culture brain glioma stem cells from clinical specimens.
ABSTRACT In recent years, the theory of brain glioma stem cells (BGSCs) has facilitated the study of gliomas. BGSCs have been accepted as the origin of gliomas and determine their biological features. Numerous efforts have been made to probe into the biological characteristics and behaviors of BGSCs. However, the culturing of target cells remains the essential first step for research on BGSCs. In this study, we established a simplified procedure to culture and isolate BGSCs from samples of clinical glioma patients. Samples of 17 glioma patients were included in the study, and the processed glioma cells were grown in serum-free stem cell media. After the tumor spheres appeared, a proliferation assay, a single-cell-derived colonies formation assay and an induced differentiation assay were carried out, followed by an immunocytochemistry assay. Serial passage was used to purify the target cells, whereas neither animal experiments nor sorting techniques were included. As a result, CD133(+) BGSCs from 8 out of 17 patients were grown and maintained in a serum-free condition combined with EGF, FGF and B-27 supplements. The tumor sphere cells were serially passaged and showed pluripotency in an induced differentiation assay. Immunocytochemistry identified the committed markers (CD133, GFAP and TU-20) and confirmed the cells were BGSCs and their progeny. The results proved that CD133(+) BGSCs from resected glioma tissue may be cultured in serum-free stem cell media, and may also be purified by conditioned culture combining serial passage, which is time-saving and cost-effective, and allows the cells to be used for subsequent research. The cell sorting techniques and animal experiments of tumorigenecity are optional. Thus, this modified procedure is more practical and feasible than other available procedures.
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ABSTRACT: Several authors have reported an increase in the incidence of brain tumors, especially among the elderly. A more complete understanding of adult glioma incidence trends might provide indications of risk factors for gliomas and contribute to the search for improved therapies. The authors used the Surveillance, Epidemiology, and End Results (SEER) registry public use data tapes, which included data on patients with cancer diagnosed between 1973 and 2000. For 3 histologies as well as for 12 histology categories combined, the authors used Poisson regression to model incidence as a function of year of diagnosis, age at diagnosis, race (white or African American), and gender. They used cubic splines to fit age at diagnosis and year of diagnosis and tested for all pair-wise interactions. The interaction between year of diagnosis and age at diagnosis was significant in all four groups modeled. In glioblastoma, there was also a significant interaction between gender and age at diagnosis. In anaplastic astrocytoma, there was a significant interaction between gender and year of diagnosis. In oligodendroglioma, there was a significant interaction between race and gender. In the 12 histology categories combined, there was a significant interaction between gender and age at diagnosis. The results in the current study were consistent with other published reports that showed an increase in the incidence of brain tumors using SEER data. Although others have observed increasing incidence trends among the elderly, the authors formally tested and found a statistically significant interaction between age at diagnosis and year of diagnosis.Cancer 12/2004; 101(10):2293-9. · 4.77 Impact Factor