gms | German Medical Science

62nd Annual Meeting of the German Society of Neurosurgery (DGNC)
Joint Meeting with the Polish Society of Neurosurgeons (PNCH)

German Society of Neurosurgery (DGNC)

7 - 11 May 2011, Hamburg

Neuroepithelial 3D cysts as a tool for studying spinal cord regeneration after injury

Meeting Abstract

  • M. Niesche - Klinik und Poliklinik für Neurochirurgie, Universitätsklinikum Carl-GHustav Carus der Technischen Universität Dresden
  • A. Meinhard - Center for Regenerative Therapies Dresden (CRTD), DFG Research Center and Cluster of Excellence, Dresden
  • G. Schackert - Klinik und Poliklinik für Neurochirurgie, Universitätsklinikum Carl-GHustav Carus der Technischen Universität Dresden
  • E. Tanaka - Center for Regenerative Therapies Dresden (CRTD), DFG Research Center and Cluster of Excellence, Dresden

Deutsche Gesellschaft für Neurochirurgie. Polnische Gesellschaft für Neurochirurgen. 62. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC), Joint Meeting mit der Polnischen Gesellschaft für Neurochirurgen (PNCH). Hamburg, 07.-11.05.2011. Düsseldorf: German Medical Science GMS Publishing House; 2011. DocDI.05.07

doi: 10.3205/11dgnc132, urn:nbn:de:0183-11dgnc1323

Published: April 28, 2011

© 2011 Niesche et al.
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Outline

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Objective: Traumatic spinal cord injury (SCI) is a devastating illness with severe disability, major associated morbidities and poor feasibility of regeneration. In this project we wanted treat organotypic cysts from murine ES cells to accomplish patterning and spatial organization resembling a developing spinal cord.

Methods: Based on the model of creating 3D neuroepithelial cysts in vitro, we used single mESC growing in a matrix. Cyst formation occurred within 5–7 days under defined conditions. The cysts resembled a developing ependymal tube at the hindbrain level. In order to shift them along the rostrocaudal axis and to induce a pattern of dorsoventral orientation as in the normal spinal cord, cysts were treated with several posteriorizing and dorsalizing/ventralizing morphogens at distinct points in time. Cyst formation, cellular development and differentiation processes were qualitatively analyzed and quantified using immunofluorescence, confocal microscopy, PCR and ISH.

Results: Embedded ESC grow clonally as cyst-like structures. Treatment with retinoic acid caudalizes cysts down to lower cervical levels compared to the developing neural axis. Caudalized cysts can be shifted along the dorsoventral axis via incubation with a specific shh-agonist to induce ventralization including the development of interneurons and motoneurons. Treatment with dorsalizing morphogens BMP and Wnt provides dorsal sensory cells. Fusing different types we generated polarized cysts with distinct dorso-ventral polarity indicating a spatial organization.

Conclusions: We developed a 3D neuroepithelial cyst culture from mESC that recapitulates spatial and temporal development and resembles the neural tube in vitro. Ependymal tube-like aggregates express embryonic antigens in defined spatial domains and react to locotypical morphogens with neuronal specification and patterning of neural tissue. Transplantable 3D aggregates should enhance regenerative ability of severed tissue, support axonal growth and restauration of the lesion. To examine the engraftment potential of the neuroepithelial cysts in vivo, we plan to implant the cysts in a rat SCI model to test tumorigenicity, axonal sprouting in various matrices, cellular and histological integration, functional improvement and maximum size of the SC lesion that can be restored.