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.

[1]  B. Maria,et al.  Targeting Hyaluronan Interactions in Malignant Gliomas and Their Drug-Resistant Multipotent Progenitors , 2008, Clinical Cancer Research.

[2]  P. Dell'Albani.  Stem Cell Markers in Gliomas , 2008, Neurochemical Research.

[3]  Daniel H. Geschwind,et al.  Cancerous stem cells can arise from pediatric brain tumors , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Deric M. Park,et al.  Biology of glioma cancer stem cells , 2009, Molecules and cells.

[5]  D. Farkas,et al.  Isolation of cancer stem cells from adult glioblastoma multiforme , 2004, Oncogene.

[6]  Bin Liu,et al.  The different HMGA1 expression of total population of glioblastoma cell line U251 and glioma stem cells isolated from U251 , 2011, Brain Research.

[7]  R. Coggeshall,et al.  Region-specific generation of cholinergic neurons from fetal human neural stem cells grafted in adult rat , 2002, Nature Neuroscience.

[8]  D. Steindler,et al.  Human cortical glial tumors contain neural stem‐like cells expressing astroglial and neuronal markers in vitro , 2002, Glia.

[9]  R. McLendon,et al.  Nonreceptor tyrosine kinase BMX maintains self-renewal and tumorigenic potential of glioblastoma stem cells by activating STAT3. , 2011, Cancer cell.

[10]  M. Andreeff,et al.  A leukemic stem cell with intrinsic drug efflux capacity in acute myeloid leukemia. , 2001, Blood.

[11]  S. Weiss,et al.  A multipotent EGF-responsive striatal embryonic progenitor cell produces neurons and astrocytes , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[12]  Tatsuya Ozawa,et al.  PTEN/PI3K/Akt pathway regulates the side population phenotype and ABCG2 activity in glioma tumor stem-like cells. , 2009, Cell stem cell.

[13]  Daniel J. Hoeppner,et al.  Distribution of CD133 reveals glioma stem cells self-renew through symmetric and asymmetric cell divisions , 2011, Cell Death and Disease.

[14]  T. Kondo,et al.  Cancer Stem Cells Persist in Many Cancer Cell Lines , 2004, Cell cycle.

[15]  G. Faccani,et al.  Glioblastoma cancer stem cells: heterogeneity, microenvironment and related therapeutic strategies , 2010, Cell biochemistry and function.

[16]  M. Souweidane,et al.  Optimization of glioblastoma multiforme stem cell isolation, transfection, and transduction , 2011, Journal of Neuro-Oncology.

[17]  Hui Wang,et al.  Turning Cancer Stem Cells Inside Out: An Exploration of Glioma Stem Cell Signaling Pathways* , 2009, The Journal of Biological Chemistry.

[18]  A. Smits,et al.  Oligodendrocyte progenitor cells can act as cell of origin for experimental glioma , 2009, Oncogene.

[19]  Kylie M. Price,et al.  Side Population is Not Necessary or Sufficient for a Cancer Stem Cell Phenotype in Glioblastoma Multiforme , 2011, Stem cells.

[20]  X. Bian,et al.  Chemoattractant receptors as pharmacological targets for elimination of glioma stem-like cells. , 2011, International immunopharmacology.

[21]  T. Kondo,et al.  Persistence of a small subpopulation of cancer stem-like cells in the C6 glioma cell line. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[22]  D. Grzybicki.  Enhanced invasion in vitro and the distribution patterns in vivo of CD133+ glioma stem cells , 2012 .

[23]  Zang Ai-hua,et al.  Stem Cells,Cancer and Cancer Stem Cells , 2005 .

[24]  R. Henkelman,et al.  Identification of human brain tumour initiating cells , 2004, Nature.

[25]  R. Campbell,et al.  Cryopreservation of cancer-initiating cells derived from glioblastoma. , 2011, Frontiers in bioscience.

[26]  Yong Yang,et al.  Isolation and characterization of cancer stem like cells in human glioblastoma cell lines. , 2009, Cancer letters.

[27]  Cynthia Hawkins,et al.  Identification of a cancer stem cell in human brain tumors. , 2003, Cancer research.

[28]  Gerald C. Chu,et al.  P53 and Pten control neural and glioma stem/progenitor cell renewal and differentiation , 2008, Nature.

[29]  Hyung-Seok Kim,et al.  The presence of stem cell marker‐expressing cells is not prognostically significant in glioblastomas , 2011, Neuropathology : official journal of the Japanese Society of Neuropathology.

[30]  W. Hall,et al.  Persistence of CD133+ cells in human and mouse glioma cell lines: detailed characterization of GL261 glioma cells with cancer stem cell-like properties. , 2008, Stem cells and development.