Research on the proliferation and drug-resistance of human BTSCs

摘要

Brain tumor stem cells (BTSC_s) have been implied to play an important role in genesis and development of glioma. However, their characteristics on proliferation and drug-resistance are uncertain thoroughly. In this experiment, some of the biological characteristics about BTSC_s were explored. Twenty cases of different grades of human glioma tissues were obtained from clinic. 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 20ng/μm of EGF, LIF and bFGF respectively. At the 5~(th) day after culture, the suspensioned cellular spheres were harvested and then both the CD133~+and CD 133~- cells were purified by immunomagnetic beads. The BTSCs were identified by testing the expression of CD133, NSE and GFAP, along with the culture process. CCK.8 was used to assay the proliferating situation of CD133~+ cells in the different grade gliomas, and to compare the drug-resistance between the CD133~+ and CD 133~ - cells in the medium containing differentrnconcentrations (including 1 Oμg/ml、20μg/ml、 40μg/ml , and 80μg/ml) of teniposide (VM-26), arnkind of drug against gliomas. The results showed that the CD133~+ cells could regenerate by self-renewal, then generate and different into NSE~+cells and GFAP~+ ones respectively. CD133~+ cells in the high grade of gliomas showed the faster generation than the ones in the low grade. The number of survived CD133~+ cells in the medium containing VM-26 (no matter any conentration) was much more than the CD133~ -.ones in it. Therefore, it was implied that the CD133~+ BTSC_s existed in the glioma tissues possessed the more tolerant ability to the VM-26, and could proliferate much more easily in the high grade glioma..