gms | German Medical Science

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

Quantum-Dot bioconjugates for the staining of living glioma cells

Meeting Abstract

  • S. Kantelhardt - Klinik für Neurochirurgie, Universitätsmedizin Göttingen
  • D. Arndt-Jovin - Max-Planck-Institut für biophysikalische Chemie, Göttingen
  • T. Jovin - Max-Planck-Institut für biophysikalische Chemie, Göttingen
  • V. Rohde - Klinik für Neurochirurgie, Universitätsmedizin Göttingen
  • A. Giese - Klinik für Neurochirurgie, Universitätsmedizin Göttingen

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. DocMI.05-10

doi: 10.3205/09dgnc201, urn:nbn:de:0183-09dgnc2015

Published: May 20, 2009

© 2009 Kantelhardt et al.
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Outline

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Objective: Recent studies have highlighted the value of complete tumor resection for patients suffering from high grade gliomas. As intraoperative differentiation of glioma cells and surrounding tissue can be very difficult, several techniques for the intraoperative identification of gliomas were proposed. Here we present a technical approach for fluorescence staining of glioma cells, which might offer the means for highly specific identification of remaining glioma tissue. We applied fluorescent nanoparticles, quantum dots (QD), coupled to targeting biomolecules.

Methods: Experiments were conducted with cell-cultures of 15 glioma cell-lines. 3 of these with defined EGFR status were applied in an animal tumor-model (2 mice for each cell line). Finally the method was applied to a human tumor biopsy, taken from a glioblastoma multiforme and brain tissue, which had to be resected during the surgical approach. Staining was performed directly after explantation under physiological conditions (isotonic solution, 37° C, native tissue samples). QD-EGF, QD coupled to 3 antibodies directed against EGFR-epitopes and QD coupled to an anti-PDGFR antibody were applied. Corresponding histologies were obtained for all samples.

Results: Specific uptake of QD-EGF into living glioma cells was found in 8 cell-lines, corresponding to the EGFR status as described in literature. As expected the other 7 cell-lines showed no uptake of QD-EGF or QD alone. Three cell lines were tested in an animal tumor-model. Again uptake corresponded to the EGFR status of the cell lines. No significant unspecific uptake was observed in all 6 mice. Tumors derived from all cell lines with over expressed EGFR and a mutation in the EGF binding site could be stained by application of QD-conjugates of antibodies that target other epitopes on the EGFR only, demonstrating the highly specific binding of QD-bioconjugates. Finally, a human glioblastoma biopsy and surrounding brain tissue were stained, resulting in highly specific binding of QD-bioconjugates. Fluorescence was sufficient to be observed macroscopically but also allowed discrimination of individual tumor cells under higher magnification.

Conclusions: We have demonstrated that QD-EGF staining is a feasible alternative for the intraoperative identification of glioma cells. QD can easily be coupled to all kinds of targeting biomolecules and therefore might offer the means for intraoperative identification of low-grade gliomas as well.