An efficient method for derivation and propagation of glioblastoma cell lines that conserves the molecular profile of their original tumours

被引:123
作者
Al-Mayhani, Talal M. Fael [1 ]
Ball, Siolian L. R. [1 ]
Zhao, Jing-Wei [1 ]
Fawcett, James [1 ]
Lchimura, Koichi [2 ]
Collins, Peter V. [2 ]
Watts, Colin [1 ,3 ]
机构
[1] Univ Cambridge, Cambridge Ctr Brain Repair, Dept Clin Neurosci, Cambridge CB2 0PY, England
[2] Univ Cambridge, Dept Mol Histopathol, Cambridge CB2 0PY, England
[3] Univ Cambridge, Dept Neurosurg, Addenbrookes Hosp, Cambridge CB2 0PY, England
关键词
Glioblastoma; Glioma; Glia; Stem cell; Progenitor; Cancer; Human; Brain; Model; Cell culture; CGH; NEURAL STEM-CELLS; HUMAN-MALIGNANT GLIOMA; ADULT HUMAN BRAIN; PROGENITOR CELLS; GROWTH; IDENTIFICATION; EXPRESSION; GENE; OLIGODENDROCYTES; DIFFERENTIATION;
D O I
10.1016/j.jneumeth.2008.07.022
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A growing body of evidence suggests that glioma stem-like cells are more representative of their parent tumours when cultured under defined serum-free conditions with the mitogens epidermal growth factor (EGF) and fibroblast growth factor (FGF). However, culturing these cells as free-floating spheroids can result in difficulty in efficiently deriving and propagating cell lines. We have combined neurosphere and monolayer culture techniques to improve the efficiency with which cells can be derived from clinical tumour samples under defined serum-free conditions. We have applied our protocol to consecutive samples of glioblastoma to show that they can form experimental tumours that recapitulate many of the histological features of the parent turnout. We go on to show that the turnout initiating cells also retain the cytogenetic abnormalities of the parent turnout. Finally we examined the cell lines for expression of markers associated with neural stem cells. Our results confirm the expression of transcription factors associated with neural patterning and specification including Sox2, Olig2, Pax6 and Nkx2.2. We went on to establish that these factors were also expressed in the parent turnout indicating that their expression was not a function of our culture conditions. The Cambridge Protocol is an efficient method of deriving stem-like tumour initiating cells from glioblastoma. improving the efficiency of derivation will facilitate the improvement of in vitro and in vivo model systems to study disease mechanisms, screen drugs and develop novel therapeutic approaches in the future. (C) 2008 Elsevier B.V. All rights reserved.
引用
收藏
页码:192 / 199
页数:8
相关论文
共 49 条
[1]   CD133+ and CD133- glioblastoma-derived cancer stem cells show differential growth characteristics and molecular profiles [J].
Beier, Dagmar ;
Hau, Peter ;
Proescholdt, Martin ;
Lohmeier, Annette ;
Wischhusen, Joerg ;
Oefner, Peter J. ;
Aigner, Ludwig ;
Brawanski, Alexander ;
Bogdahn, Ulrich ;
Beier, Christoph P. .
CANCER RESEARCH, 2007, 67 (09) :4010-4015
[2]   CYTOGENETICS OF HUMAN BRAIN-TUMORS [J].
BIGNER, SH ;
MARK, J ;
BIGNER, DD .
CANCER GENETICS AND CYTOGENETICS, 1990, 47 (02) :141-154
[3]   CHROMOSOMAL PROGRESSION OF MALIGNANT HUMAN GLIOMAS FROM BIOPSY TO ESTABLISHMENT AS PERMANENT LINES INVITRO [J].
BIGNER, SH ;
MARK, J ;
BIGNER, DD .
CANCER GENETICS AND CYTOGENETICS, 1987, 24 (01) :163-176
[4]  
BIGNER SH, 1990, MONOGR PATHOL, V32, P30
[5]   Shared oligodendrocyte lineage gene expression in gliomas and oligodendrocyte progenitor cells [J].
Bouvier, C ;
Bartoli, C ;
Aguirre-Cruz, L ;
Virard, I ;
Colin, C ;
Fernandez, C ;
Gouvernet, J ;
Figarella-Branger, D .
JOURNAL OF NEUROSURGERY, 2003, 99 (02) :344-350
[6]  
CARLSSON J, 1978, ACTA PATH MICRO IM A, V86, P45
[7]   Revisiting the astrocyte-oligodendrocyte relationship in the adult CNS [J].
Carmen, Jessica ;
Magnus, Tim ;
Cassiani-Ingoni, Riccardo ;
Sherman, Larry ;
Rao, Mahendra S. ;
Mattson, Mark R. .
PROGRESS IN NEUROBIOLOGY, 2007, 82 (03) :151-162
[8]   Molecular analysis of microdissected de novo glioblastomas and paired astrocytic tumors [J].
Cheng, Y ;
Ng, HK ;
Ding, M ;
Zhang, SF ;
Pang, JCS ;
Lo, KW .
JOURNAL OF NEUROPATHOLOGY AND EXPERIMENTAL NEUROLOGY, 1999, 58 (02) :120-128
[9]   FINE-STRUCTURAL STUDIES ON CULTURED HUMAN GLIAL AND GLIOMA-CELLS - TECHNIQUES AND APPLICATIONS [J].
COLLINS, VP ;
BRUNK, UT .
ULTRASTRUCTURAL PATHOLOGY, 1984, 7 (04) :321-334
[10]   Niche-independent symmetrical self-renewal of a mammalian tissue stem cell [J].
Conti, L ;
Pollard, SM ;
Gorba, T ;
Reitano, E ;
Toselli, M ;
Biella, G ;
Sun, YR ;
Sanzone, S ;
Ying, QL ;
Cattaneo, E ;
Smith, A .
PLOS BIOLOGY, 2005, 3 (09) :1594-1606