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73. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC)
Joint Meeting mit der Griechischen Gesellschaft für Neurochirurgie

Deutsche Gesellschaft für Neurochirurgie (DGNC) e. V.

29.05. - 01.06.2022, Köln

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IL-17A, IL-22, IL-23 and CCL20 – influence on migration and invasion of rat glioblastoma cell lines using in vitro techniques and a rat organotypic brain slice co-culture (OBSC)

IL-17A, IL-22, IL-23 und CCL20 – Einfluss auf Migration und Invasion von Ratten Glioblastomzelllinien unter Verwendung von in vitro Methoden und organotypischen Rattenhirnschnitt-Co-Kulturen (OBSC)

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  • presenting/speaker Johannes Falter - Universitätsklinikum Regensburg, Klinik und Poliklinik für Neurochirurgie, Regensburg, Deutschland
  • Anette Lohmeier - Universitätsklinikum Regensburg, Klinik und Poliklinik für Neurochirurgie, Regensburg, Deutschland
  • Petra Eberl - Universitätsklinikum Regensburg, Klinik und Poliklinik für Neurochirurgie, Regensburg, Deutschland
  • Eva-Maria Störr - Universitätsklinikum Regensburg, Klinik und Poliklinik für Neurochirurgie, Regensburg, Deutschland
  • Nils-Ole Schmidt - Universitätsklinikum Regensburg, Klinik und Poliklinik für Neurochirurgie, Regensburg, Deutschland
  • Martin Proescholdt - Universitätsklinikum Regensburg, Klinik und Poliklinik für Neurochirurgie, Regensburg, Deutschland

Deutsche Gesellschaft für Neurochirurgie. 73. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC), Joint Meeting mit der Griechischen Gesellschaft für Neurochirurgie. Köln, 29.05.-01.06.2022. Düsseldorf: German Medical Science GMS Publishing House; 2022. DocP152

doi: 10.3205/22dgnc465, urn:nbn:de:0183-22dgnc4656

Published: May 25, 2022

© 2022 Falter 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

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Objective: Tumor cell invasion has been demonstrated to be enhanced by inflammatory cytokines such as IL-17A, IL-22, IL-23 and CCL20 in various tumor entities other than glioblastoma (GBM). We have tested the impact of these factors on GBM migration and invasion utilizing four different in vitro techniques.

Methods: For avoiding xenogenous effects in the rat organotypic brain slice co-culture (OBSC), we use the rat glioblastoma cell lines 9L and S635 and as a non-neoplastic control cell line we used primary rat astrocytes. We employed a Boyden Chamber Invasion assay (transwell with matrigel), spheroid migration assay (4000 cell spheroid in round bottom well) and a wound healing migration assay. Moreover we have adopted and evolved rat OBSC to test for pro-migratory effects of the candidate cytokines: we use 6-12d rat brains, cut them in 350µm slices, put them on a 0.4µm insert swimming on culture medium. Next, we placed 4000 PKH67 fluorescence-labeled rat glioblastoma cells as spheroid-like structures in the area of the hippocampus next to the lateral ventricle and quantified the migration for 7 days.

Results: IL-22 shows enhancement of invasion and migration in our tested glioblastoma cell lines but not in the normal astrocytes. IL-17A shows enhancement of migration only on 9L cell lines. The establishment of the rat OBSC was feasible in our set-up and viability of the co-culture was shown over the course of 7 days, yet the effect of the above mentioned factors was not reproducible in the OBSC.

Conclusion: The effect of enhanced migration and/or invasion was not reproducible with our rat OBSC, probably because of minor discriminatory power due to our method of measuring the 2D extent of fluorescence. Refinement of measurement techniques might lead to more conclusive data in our rat OBSC. The downstream pathways leading to the effect with IL-22 and the effect on human cell lines remain to be examined.