Article
Stem-like glioma cells comprise a non-homogeneous population of cells with varying degrees of stemness and tumorigenicity
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Authors
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Darius Kalasauskas
- The Translational Neurooncology Research Group, Department of Neurosurgery, University Medical Centre, Johannes Gutenberg University, Mainz, Germany
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Anne Rupp
- The Translational Neurooncology Research Group, Department of Neurosurgery, University Medical Centre, Johannes Gutenberg University, Mainz, Germany
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Mirjam Renovanz
- The Translational Neurooncology Research Group, Department of Neurosurgery, University Medical Centre, Johannes Gutenberg University, Mainz, Germany
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Bettina Sprang
- The Translational Neurooncology Research Group, Department of Neurosurgery, University Medical Centre, Johannes Gutenberg University, Mainz, Germany
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Marina Janocha
- The Translational Neurooncology Research Group, Department of Neurosurgery, University Medical Centre, Johannes Gutenberg University, Mainz, Germany
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Walter Schulz-Schaeffer
- Department of Neuropathology, University Medical Centre Göttingen, Göttingen, Germany
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Ella L. Kim
- The Translational Neurooncology Research Group, Department of Neurosurgery, University Medical Centre, Johannes Gutenberg University, Mainz, Germany
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Alf Giese
- The Translational Neurooncology Research Group, Department of Neurosurgery, University Medical Centre, Johannes Gutenberg University, Mainz, Germany
| Published: | May 21, 2013 |
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Outline
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Objective: To investigate the relation between key attributes of stemness, self-renewal and differentiation, and tumorigenicity in stem-like glioma cells derived from human glioblastoma multiforme.
Method: Brain Tumor Initiating Cells (BTIC) were derived from glioma cell lines or fresh glioblastoma specimens. Neurosphere cultures were maintained in Neurobasal medium supplemented with bFGF and EGF (10 and 20 ng/ml, respectively). For immunofluorescence staining, single cell suspensions were plated onto ornithine coated cover slips and incubated to allow cells to adhere. BrdU staining and detection was performed by using BrdU (5-bromo-2'-deoxyuridine) detection kit (Roche). For intracranial implantation, single cell suspension was injected into the caudato-putamen of the right-brain hemisphere of immunodefficient nude mice. Mice were observed daily and sacrified upon manifestation of neurological symptoms. 1–3 μm thick sections were prepared and stained with the indicated antibodies.
Results: 7 BTIC cell lines were tested. All of them were able to self-renew, induce tumors in vivo and expressed constitutively nestin, a marker of neural stem cells. However, under differentiation inducing conditions only 3 of these cell lines showed the ability to undergo phenotypic differentiation. Strikingly, differentiation-capable cell lines undergo a marked cessation in the proliferative capacity as evidenced by the drop of BrdU incorporation rate under differentiation inducing conditions. Contrary, differentiation-incapable cell lines are either incapable or considerably less capable of the proliferative inhibition under differentiation-inducing conditions. The ability or inability to undergo proliferative block under differentiation-inducing conditions in vitro correlates with the degree of tumorigenicity as revealed in in vivo experiments using an orthotopic glioma model. Parallel assessment of the proliferative and tumorigenic potentials in vitro and in vivo, respectively, shows that the rate of growth of tumours initiated by differentiation-capable glioma cells is significantly longer compared to that of differentiation-incapable glioma cells.
Conclusions: 1. BTICs comprise a heterogeneous population of cells differing not only phenotypically but also with respect to clinicopathological properties. 2. Our data do not support the postulate that the ability to differentiate is a criterion for identification of the most tumorigenic types of glioma cells.
