gms | German Medical Science

57th Annual Meeting of the German Society of Neurosurgery
Joint Meeting with the Japanese Neurosurgical Society

German Society of Neurosurgery (DGNC)

11 - 14 May, Essen

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Induction of differentiation of murine neural stem cells by confrontation with highly regenerative blastema tissue of the axolotl salamander

Induzierung von Differenzierungsmechanismen von murinen neuralen Stammzellen durch Konfrontation von regenerierendem Blastemgewebe des Axolotlsalamanders

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  • corresponding author M. Kirsch - Klinik und Poliklinik für Neurochirurgie, Carl Gustav Carus Universitätsklinikum, Technische Universität Dresden
  • M. Niesche - Klinik und Poliklinik für Neurochirurgie, Carl Gustav Carus Universitätsklinikum, Technische Universität Dresden
  • B. Kilian - Klinik und Poliklinik für Neurochirurgie, Carl Gustav Carus Universitätsklinikum, Technische Universität Dresden
  • G. Schackert - Klinik und Poliklinik für Neurochirurgie, Carl Gustav Carus Universitätsklinikum, Technische Universität Dresden
  • H.K. Schackert - Chirurgische Forschung, Carl Gustav Carus Universitätsklinikum, Technische Universität Dresden
  • E. Tanaka - Max Planck Institute for Molecular Cell Biology and Genetics, Dresden

Deutsche Gesellschaft für Neurochirurgie. Japanische Gesellschaft für Neurochirurgie. 57. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e.V. (DGNC), Joint Meeting mit der Japanischen Gesellschaft für Neurochirurgie. Essen, 11.-14.05.2006. Düsseldorf, Köln: German Medical Science; 2006. DocSO.03.08

The electronic version of this article is the complete one and can be found online at: http://www.egms.de/en/meetings/dgnc2006/06dgnc180.shtml

Published: May 8, 2006

© 2006 Kirsch 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: Caudate amphibians such as the Axolotl salamander (Ambystoma mexicanum) regenerate an amputed tail including a fully functional spinal cord. After amputation, a multipotent array of proliferating and differentiating cells termed blastema develops which gives rise to all cell types of the regenerating tail and spinal cord. This model represents an opportunity to understand what factors are required to induce a neural stem cell to undergo productive regeneration. The purpose of our study was to investigate the differentiation potential of murine neural stem cells (NSC) in response to intrinsic signalling of the blastema.

Methods: Primary murine embryonic cerebellar neural stem cells were grown as neurospheres in neural stem cell medium. Blastema were obtained 5-7 days after amputation of the axolotl tail. After removal of the overlaying skin, the blastema was removed under a dissecting microscope. Murine neurospheres were juxtaposed with blastema tissue in vitro. The invasion of NSC into the blastema was analyzed by confocal microscopy. Differentiation and proliferation of NSC was analysed using specific antibodies against Nestin, NeuN, β3-Tubulin, GFAP, Ki67, PCNA, BrdU, CNPase, and GalC.

Results: In vitro conditions were optimized to allow both NSC and blastema cells to survive and proliferate. Within 3 days NSC were able to invade into the regenerating blastema. Depending on their location in relation to the blastema, proliferation occurred mainly in the center of the neurosphere contrasting to isolated neurospheres which show predominant proliferation in the periphery. Upon confrontation, a strong differentiation response was seen showing an increase of oligodendroglial and neuronal cells but a decrease of precursor and astrocytic cells as compared to conventional in vitro induction of differentiation via growth factor withdrawal.

Conclusions: Murine NSC responded to a regenerative environment such as axolotl blastema tissue and revealed a higher differentiation potential compared to standard induction of differentiation. Further studies are aiming to identify the factors responsible for the higher regenerative potential induced by blastema tissue.