Isolation, cultivation and identification of brain glioma stem cells by magnetic bead sorting.
This study describes a detailed process for obtaining brain glioma stem cells from freshly dissected human brain glioma samples using an immunomagnetic bead technique combined with serum-free media pressure screening. Furthermore, the proliferation, differentiation and self-renewal biological features of brain glioma stem cells were identified. Results showed that a small number of CD133 positive tumor cells isolated from brain glioma samples survived as a cell suspension in serum-free media and proliferated. Subcultured CD133 positive cells maintained a potent self-renewal and proliferative ability, and expressed the stem cell-specific markers CD133 and nestin. After incubation with fetal bovine serum, the number of glial fibrillary acidic protein and microtubule associated protein 2 positive cells increased significantly, indicating that the cultured brain glioma stem cells can differentiate into astrocytes and neurons. Western blot analysis showed that tumor suppressor phosphatase and tensin homolog was highly expressed in tumor spheres compared with the differentiated tumor cells. These experimental findings indicate that the immunomagnetic beads technique is a useful method to obtain brain glioma stem cells from human brain tumors.
Department of Neurosurgery, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221000, Jiangsu Province, China ; Laboratory of Neurosurgery, Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China ; Jiangsu Key Laboratory of Brain Disease Biology, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221000, Jiangsu Province, China.
Xiuping
X
Zhou
Graduate School, Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China.
Chao
C
Zheng
Department of Neurosurgery, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221000, Jiangsu Province, China ; Laboratory of Neurosurgery, Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China ; Jiangsu Key Laboratory of Brain Disease Biology, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221000, Jiangsu Province, China.
Qiong
Q
Shi
Department of Neurosurgery, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221000, Jiangsu Province, China.
Xiang
X
Li
Department of Neurosurgery, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221000, Jiangsu Province, China.
Zhigang
Z
Shen
Department of Neurosurgery, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221000, Jiangsu Province, China ; Laboratory of Neurosurgery, Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China ; Jiangsu Key Laboratory of Brain Disease Biology, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221000, Jiangsu Province, China.
Rutong
R
Yu
CD133; brain glioma stem cells; glial fibrillary acidic protein; immunomagnetic beads; microtubule associated protein 2; nestin; neural regeneration