gms | German Medical Science

63rd Annual Meeting of the German Society of Neurosurgery (DGNC)
Joint Meeting with the Japanese Neurosurgical Society (JNS)

German Society of Neurosurgery (DGNC)

13 - 16 June 2012, Leipzig

In vitro 3D reconstitution of the mammalian spinal cord using murine embryonic stem cells

Meeting Abstract

  • M. Niesche - Klinik und Poliklinik für Neurochirurgie, Universitätsklinikum der TU Dresden
  • A. Meinhardt - Zentrum für Regenerative Therapien (CRTD) der TU Dresden
  • E. Tanaka - Zentrum für Regenerative Therapien (CRTD) der TU Dresden
  • M. Kirsch - Klinik und Poliklinik für Neurochirurgie, Universitätsklinikum der TU Dresden
  • G. Schackert - Klinik und Poliklinik für Neurochirurgie, Universitätsklinikum der TU Dresden

Deutsche Gesellschaft für Neurochirurgie. Japanische Gesellschaft für Neurochirurgie. 63. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC), Joint Meeting mit der Japanischen Gesellschaft für Neurochirurgie (JNS). Leipzig, 13.-16.06.2012. Düsseldorf: German Medical Science GMS Publishing House; 2012. DocDO.08.12

doi: 10.3205/12dgnc078, urn:nbn:de:0183-12dgnc0781

Published: June 4, 2012

© 2012 Niesche et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.



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 3D neuroepithelial tubes from murine ES cells to accomplish patterning and spatial organization resembling a developing spinal cord.

Methods: Based on the model of creating 3D cysts in vitro we used single mESC growing in a matrix. Tube formation occurred within 5–7 days under defined conditions. They resemble 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, tubes were treated with several posteriorizing and dorsalizing/ventralizing morphogens at distinct time points. Tube formation, cellular development and differentiation processes were qualitatively analysed and quantified using immunofluorescence, confocal microscopy, PCR and ISH.

Results: Embedded ESC grow clonally as tube like structures. They possess apical-basal polarity, express neural stem cell markers and differentiate along the neural lineage, following normal embryonic development. Treatment with EC23 caudalizes them down to lower cervical levels of the developing neural axis. Caudalized tubes can be shifted along the dorsoventral axis. Incubation with an agonist of the shh-pathway induces ventralization including the development of interneurons and motorneurons. Treatment with dorsalizing morphogens BMP7 and Wnt1 provides dorsal sensory progenitors. In vitro fusion of both types we generated polarized tubes with distinct dorso-ventral polarity indicating a spatial organization.

Conclusions: We developed a 3D neuroepithelial tube 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 restoration of the lesion. To examine the engraftment potential of the neuroepithelial tubes in vivo, we plan to implant the tubes in a rodent SCI model to test tumorigenicity, axonal sprouting, cellular and histological integration, functional improvement and restoration potential of the SCI lesion.