The Different Radio Sensitivity Between Normal Glioma Cells And Brain Tumor Stem Cells

Objective:1.Isolation,culture and identifieation of brain tumor stem cells(BTSCs) in vitro. Observe the grow pattern of BTSCs.2.To explore The different radio sensitivity between normal glioma cells and brain tumor stem cells.Methods:1.The tumor from Patient sunder going tumor resections were acutely dissociated, triturate into single cell and then filter, Gliomacell line was seeded into DMEM/F12medium. The brain tumor stem cells were sorted from U251cell line by means of serum starvation of serum-free DMEM/F12medium supplemented withbFGF、EGF、B27.After the brain tumor spheres generated, Partlythey were separated in free-medium. Monoclonal culture were Progressed simultaneously. Partly Brain tumors spheres were seeded in serum DMEM/F12medium, then the differentiation ability was detected.2.After glioma cells and BTSCs were irradiated by0、2、4、6、8、10Gy X-ray,and two kind of cells were continuously cultured36h, the ratios of CD133+cells were checked with Flow cytometry. Furthermore, the proliferation activity of glioma cells and BTSCs were tested by MTT. The radiosensitivity of glioma cells and BTSCs were measured by clone forming assay.Results:1. Only a few tumor cells survived in serum-free medium And suspendedly Proliferated into the cells sphere. The cells spheres till kept the capability of self-renewingandProliferation and Presenting CD133PositiveProduets. The ratio of CD133+cell in BTSCs were increased with the increase of irradiate dose. But it no obviously changed in normal glioma cells. 2. After glioma cells and BTSCs were irradiated by0、2、4、6、8、10GYX-ray,The survival rates of two kind of cells were both decreased, while more obvious decrease can be seen in glioma cells than BTSCs.(P＜0.05).3. The clone forming rate of glioma cells and BTSCs were decreased. But BTSCs was higher than normal glioma cells after receiving irradiation.(P＜0.05).Conclusion1. Within human glioblastoma there were brain tumor stem cells, which has the same properties with normal neural stem cells. CD133is key surface markers for BTSCs.2. The BTSCs’radiosensitivity is obviously lower than that of glioma cells.