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Updated: Jul 12, 2026
Processing of Primary Brain Tumor Tissue for Stem Cell Assays and Flow Sorting
Published on: September 25, 2012
[Chemoresistance of CD133(+) tumor stem cells from human brain glioma]
Chang-Long Bi1, Jia-Sheng Fang, Feng-Hua Chen
1Department of Neurosurgery, Xiangya Hospital, Central South University, Changsha 410008, China. 1975310@sina.com.cn
Objective:
To explore the multidrug resistance (MDR) mechanism of ABC superfamily transporters in the tumor stem cells(TSC) from human brain glioma tissues.
Methods:
Samples of glioma were obtained from 30 patients undergoing microsurgical tumor resection. The CD133(+) cells and CD133(-) cells from these tumor specimens were isolated by magnetic activated cell sorting(MACS). These cells were cultured, proliferated and passaged. The protein and activity expression of multidrug-resistance protein 1(MDR1) and multidrug-resistance associated protein 1(MRP1) were analyzed between CD133(+) and CD133(-) cells by immunocytochemistry and RT-PCR respectively.
Results:
CD133(+) cells generated free floating neurosphere like brain tumor spheres(BTS) and abnormal proliferating capacity in the serum-free medium(SFM) in vitro. Three cases from glioblastoma stem cells could form BTS in the complete medium, and could be cultured for 1-3 passages. The range of positive cell proportion for MDR1 and MRP1 expression in CD133(+) cells was 18%-67% and 23%-73% respectively. The expression levels of MDR1 and MRP1 mRNA were higher in CD133(+) glioma stem cells than those in the differentiated tumor cells(TC), the protein activity was increased to 16.1 and 19.6 times respectively compared with that of TC. The protein and activity expression were positively related to the pathological grades of tumors. MDR1 or MRP1 drug resistance was not expressed in all the tumors and there was obvious correlation between MDR1 and MRP1.
Conclusion:
Only a small proportion of cells in the heterogeneous glioma is CD133(+) brain tumor stem cells which display the strong capacity of self-renewing, abnormal proliferation and intrinsic multidrug resistance to traditional chemotherapy. The high expression of MDR1 and MRP1 by the CD133(+) brain tumor stem cells is one of the main mechanisms in the chemoresistance of tumors. CD133(+) brain tumor stem cells can be served as the root of multidrug resistance and key therapeutic target for glioma chemotherapy.
Insights
Glioma stem cells (CD133+) exhibit multidrug resistance (MDR) due to high expression of MDR1 and MRP1. These cells are key targets for improving glioma chemotherapy.
Area of Science:
- Neuroscience
- Oncology
- Molecular Biology
Background:
- Glioblastoma is a heterogeneous tumor with a subpopulation of cancer stem cells (CSCs).
- Multidrug resistance (MDR) in CSCs poses a significant challenge in effective cancer chemotherapy.
- Understanding the mechanisms of MDR in glioma CSCs is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate the role of ABC superfamily transporters in the MDR of human brain glioma tumor stem cells (TSCs).
- To analyze the expression and activity of MDR1 and MRP1 in CD133+ and CD133- glioma cells.
Main Methods:
- Isolation of CD133+ and CD133- cells from 30 glioma patient samples using magnetic activated cell sorting (MACS).
- Culture and proliferation of isolated cells in vitro.
- Analysis of multidrug-resistance protein 1 (MDR1) and multidrug-resistance associated protein 1 (MRP1) expression and activity using immunocytochemistry and RT-PCR.
Main Results:
- CD133+ cells demonstrated self-renewal and abnormal proliferation, forming brain tumor spheres (BTS).
- Higher expression of MDR1 and MRP1 mRNA and protein activity was observed in CD133+ glioma stem cells compared to differentiated tumor cells.
- MDR1 and MRP1 expression levels correlated positively with glioma pathological grades.
Conclusions:
- CD133+ brain tumor stem cells possess intrinsic multidrug resistance, contributing to tumor chemoresistance.
- High expression of MDR1 and MRP1 in CD133+ cells is a primary mechanism of chemoresistance in gliomas.
- Targeting CD133+ brain tumor stem cells offers a promising strategy for overcoming multidrug resistance in glioma chemotherapy.
