gms | German Medical Science

58. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e. V. (DGNC)

Deutsche Gesellschaft für Neurochirurgie (DGNC) e. V.

26. bis 29.04.2007, Leipzig

Stem cell characteristics in established glioblastoma cell lines

Stammzell-Eigenschaften in etablierten Glioblastom-Zelllinien

Meeting Abstract

  • corresponding author C. Dictus - Abteilung für Neurochirurgische Forschung, Klinik für Neurochirurgie, Universitätsklinikum Heidelberg
  • H. Mattern - Abteilung für Neurochirurgische Forschung, Klinik für Neurochirurgie, Universitätsklinikum Heidelberg
  • B. Radlwimmer - Abteilung Molekulare Genetik, DKFZ Heidelberg
  • A. Unterberg - Abteilung für Neurochirurgische Forschung, Klinik für Neurochirurgie, Universitätsklinikum Heidelberg
  • C. Herold-Mende - Abteilung für Neurochirurgische Forschung, Klinik für Neurochirurgie, Universitätsklinikum Heidelberg

Deutsche Gesellschaft für Neurochirurgie. 58. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e.V. (DGNC). Leipzig, 26.-29.04.2007. Düsseldorf: German Medical Science GMS Publishing House; 2007. DocFR.09.06

Die elektronische Version dieses Artikels ist vollständig und ist verfügbar unter: http://www.egms.de/de/meetings/dgnc2007/07dgnc127.shtml

Veröffentlicht: 11. April 2007

© 2007 Dictus et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielf&aauml;ltigt, verbreitet und &oauml;ffentlich zug&aauml;nglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Gliederung

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Objective: Recently, tumor stem cells (TSCs) have been isolated from various tumors including gliomas. These glioma-derived TSCs share features known from neural stem cells including self-renewal capacity, multipotency and spheroid formation, are highly tumorigenic and can be distinguished from other tumor cells by the expression of neural stem cell markers. So far, they have been isolated from freshly dissected tumor specimens. In the present study, we aimed to determine stem cell characteristics in tumor cells derived from established glioblastoma (GBM) cell lines.

Methods: Two GBM cell lines (NCH82, NCH89) were established as adherent cell cultures in medium supplemented with fetal calf serum (FCS) and genetically characterized by matrix CGH. These cell lines then (1) were cultured in FCS medium or stem cell-optimized (SC) medium, (2) were separated into CD133+ and CD133- fractions by magnetic beads (MB) selection, and (3) were differentiated. For all conditions, these cell lines were characterized by their expression of stem cell and differentiation markers (immunofluorescence, qRT-PCR) and their clonal expansion (limited dilution assay).

Results: Both cell lines displayed genetic profiles typical of GBMs. When cultured in SC medium, they grew as spheroids and expressed the stem cell markers CD133, nestin, Musashi-1, Sox-2 and Oct3/4; when cultured in FCS medium, they grew as adherent monolayers and downregulated their stem cell marker expression. Self-renewal capacity was confirmed by limited dilution assays where more than 50% of cells grown in SC medium were able to form spheroids from a single tumor cell. After CD133-targeted MB selection, every fraction independent of CD133 expression status grew as spheroids in SC medium and as adherent monolayers in FCS medium. After differentiation with RA/cAMP, expression of stem cell markers was downregulated in both cell lines while differentiation markers (GFAP, β III tubulin) were uregulated in NCH82 und largely unaffected in NCH89. Furthermore, self renewal capacity decreased significantly in differentiated cells.

Conclusions: In the present study, we demonstrated that TSC characteristics are maintained in a subpopulation of cells in two established GBM cell lines. After differentiation, cells lost their TSC characteristics suggesting that in vivo differentiation might be a promising future therapeutic strategy. However, the tumorigenic potential of these cells still needs to be evaluated.