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60th Annual Meeting of the German Society of Neurosurgery (DGNC)
Joint Meeting with the Benelux countries and Bulgaria

German Society of Neurosurgery (DGNC)

24 - 27 May 2009, Münster

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Mesenchymal stem cells contribute to glioblastoma: a new protocol for isolation emanating from the integrative analysis of publicly available gene expression data

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  • V. Albrecht - Tumorbiologisches Labor, Neurochirurgische Klinik, Ludwig-Maximilians-Universität München
  • K.T. Guo - Tumorbiologisches Labor, Neurochirurgische Klinik, Ludwig-Maximilians-Universität München
  • K. Juerchott - Institut für Biologie und Biochemie, AG Bioinformatik, Universität Potsdam
  • R. Goldbrunner - Tumorbiologisches Labor, Neurochirurgische Klinik, Ludwig-Maximilians-Universität München
  • J.-C. Tonn - Tumorbiologisches Labor, Neurochirurgische Klinik, Ludwig-Maximilians-Universität München
  • C. Schichor - Tumorbiologisches Labor, Neurochirurgische Klinik, Ludwig-Maximilians-Universität München

Deutsche Gesellschaft für Neurochirurgie. 60. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC), Joint Meeting mit den Benelux-Ländern und Bulgarien. Münster, 24.-27.05.2009. Düsseldorf: German Medical Science GMS Publishing House; 2009. DocP06-05

doi: 10.3205/09dgnc309, urn:nbn:de:0183-09dgnc3097

Published: May 20, 2009

© 2009 Albrecht et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en). You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.

Outline

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Objective: Malignant gliomas have been described as heterogeneous tumors which recruit a broad variety of host precursor cells. Analyzing publicly available array data for glioblastoma, brain and mesenchymal stem cells (MSCs), we identified high positive correlations in gene expression for glioblastoma and bone marrow derived mesenchymal stromal cells (bmMSCs). Thus, it was emphasized that hMSCs are incorporated in glioma tissue. Purification and characterization of MSCs from glioblastoma tissue (gbMSCs) are prerequisites for further analysis. Therefore, we established a new protocol for isolation and purification of gbMSCs from glioblastoma tissue.

Methods: Selected publicly available expression data generated in individual experiments with normal brain, glioblastoma (GBM) and bmMSC material on Affymetrix hgu133plus2 arrays were integrated in a combined expression matrix. The correlations of the rank-transformed expression profiles of the three groups were analyzed. Two independent selections of probe sets specific for bmMSC were investigated with respect to their expression in GBM. For isolation of gbMSCs, a new protocol was developed, using a gradient based isolation technique combined with magnetic bead negative sorting. The isolated cells (gbMSCs) were characterized using extensive FACS analysis. Multipotency of the cells was confirmed via differentiation assays into mesenchymal lineages.

Results: Correlations between glioblastoma samples and bmMSC samples were significantly higher than the correlation between the normal brain group and the bmMSC samples. 66 probe sets with high expression in bmMSC and low expression in brain were identified. The majority of probes (67%) showed significantly higher expression in glioblastoma tissue compared to normal brain. A second set of genes based on published MSC markers also confirmed a clear tendency towards increased expression in GBM compared to normal brain. Flow cytometry analysis of gbMSCs displayed close similarities to bmMSCs. Differentiation assays showed the osteogenic and adipogenic differentiation potential of gbMSCs, thereby displaying multipotency.

Conclusions: Our new approach of comparative analysis of publicly available expression datasets showed close similarities between mesenchymal stem cells and glioblastoma tissue. Demonstrating the presence of gbMSCs in malignant gliomas, we established a new protocol for isolation and expansion of mesenchymal stem cells from tumor tissue.