Identification of brain tumor stem cells and research on their characters of proliferation and drug-resistance / 中华神经医学杂志

ABSTRACT

Objective To isolate and culture brain tumor stem cells (BTSCs) from glioma tissues and explore the biological characteristics of BTSCs. Methods Different grade glioma tissues were obtained from 20 clinical cases. After tumors were dissociated, the sample was triturated into the single cells and then filtered. The primary glioma cells were collected and cultured in the DMEM/F12 medium containing epidermal growth factor (EGF), leukemia inhibitory factor (LIF) and basic fibroblast growth factor (bFGF), in order to promote the proliferation of BTSCs. CD133 + cells were separated by immunomagnetic bead method and identified by testing the expressions of CD133, NSE and GFAP using immunocytochemistry. CCK8 was employed to assay the proliferating situation of CD133+ cells in the different grade gliomas, and to compare the drug resistance between the CD133+ and CD133- cells in the medium containing VM-26. Results CD133+ cells were successfully separated from glioma tissues.CD133+ cells proliferated by self-renewal, then differentiated into NSE+ cells and GFAP+ ones respectively. CD133+ cells in the high grade gliomas showed the faster generation than the ones in the low grade gliomas. CD133+ cells survived more easily than the CD133- cells in the medium containing VM-26. Conclusions BTSCs exist in the glioma tissues, and possess the more tolerant to the VM-26.CD133+ cells in the high grade glioma can proliferate much more easily.