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68th Annual Meeting of the German Society of Neurosurgery (DGNC)
7th Joint Meeting with the British Neurosurgical Society (SBNS)

German Society of Neurosurgery (DGNC)

14 - 17 May 2017, Magdeburg

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Monitoring cell death and immune cell interactions in Glioblastoma with IncuCyteZOOM

Meeting Abstract

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  • Haouraa Mostafa - Division of Experimental Anesthesiology, University Hospital Ulm, Ulm, Deutschland
  • Elvira Dietrich - Division of Experimental Anesthesiology, University Hospital Ulm, Ulm, Deutschland
  • Philip Sander - Division of Experimental Anesthesiology, University Hospital Ulm, Ulm, Deutschland
  • Ayman Soboh - Division of Experimental Anesthesiology, University Hospital Ulm, Ulm, Deutschland
  • Andrej Pala - Dept. of Neurosurgery, Bezirkskrankenhaus Günzburg, Günzburg, Deutschland
  • Angelika Scheuerle - Division of Neuropathology, Bezirkskrankenhaus Günzburg, Günzburg, Deutschland
  • C. Rainer Wirtz - Dept. of Neurosurgery, Bezirkskrankenhaus Günzburg, Günzburg, Deutschland
  • Michael Georgieff - Dept. of Anesthesiology, University Hospital Ulm, Ulm, Deutschland
  • E. Marion Schneider - Division of Experimental Anesthesiology, University Hospital Ulm, Ulm, Deutschland

Deutsche Gesellschaft für Neurochirurgie. Society of British Neurological Surgeons. 68. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC), 7. Joint Meeting mit der Society of British Neurological Surgeons (SBNS). Magdeburg, 14.-17.05.2017. Düsseldorf: German Medical Science GMS Publishing House; 2017. DocMi.28.04

doi: 10.3205/17dgnc561, urn:nbn:de:0183-17dgnc5617

Published: June 9, 2017

© 2017 Mostafa et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 License. See license information at http://creativecommons.org/licenses/by/4.0/.

Outline

Text

Objective: In an attempt to test allogeneic and autologous immune sensitization and vulnerability of established and new chemotherapeutic drugs, we established permanent cell lines from patients with glioblastoma multiforme (GBM).

Methods: Fresh tumor tissue was minced and trypsinized, then subjected to Ficoll-separation and culture using endotoxin-free fetal calf serum and Iscove’s modified essential medium. Permanent cell lines were established 4-8 weeks after culture initiation. Blood-derived allogeneic and autologous mononuclear cells were activated with Interleukin- (IL-) 2 and IL-15 using standard protocols. GBM derived cell lines were characterized using flow cytometry and stem cell contents were determined by CD133 staining. For quantification of proliferation, cell death and the interaction with immune cells, we used the IncuCyteZOOM incubator microscope and software. Cells were seeded into 96well (flat-bottomed) microtiter plates and caspase 3/, Annexin-V and PI-staining were used for data analysis.

Results: Following IL-2/IL-15 activation protocols, we found active cell lysis of GBM cell lines in the presence of IL-2 alone. However, microscopical analysis demonstrated the renewal of malignant clones much faster than effective cell killing. Microscopical examination further demonstrated massive diminution of co-cultured T- and NK lymphocytes by a process likely to be due to macropinocytosis.

Conclusion: A novel mechanism identified in glioblastoma tumor cells appears to be due to macropinocytosis of immune effectors even in the presence of T- and NK cell growth factor.