gms | German Medical Science

62. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC)
Joint Meeting mit der Polnischen Gesellschaft für Neurochirurgen (PNCH)

Deutsche Gesellschaft für Neurochirurgie (DGNC) e. V.

07. - 11. Mai 2011, Hamburg

The spinal cord – motor cortex co-culture model: inducing cortical outgrowth

Meeting Abstract

Suche in Medline nach

  • J. Glumm - Institute of Cell Biology and Neurobiology, Center for Anatomy, Charité - Universitätsmedizin Berlin; Department of Neurosurgery, HELIOS Klinikum Berlin, Klinikum Buch
  • M. Pohland - Institute of Cell Biology and Neurobiology, Center for Anatomy, Charité - Universitätsmedizin Berlin
  • J. Kiwit - Department of Neurosurgery, HELIOS Klinikum Berlin, Klinikum Buch

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. DocP 081

DOI: 10.3205/11dgnc302, URN: urn:nbn:de:0183-11dgnc3029

Veröffentlicht: 28. April 2011

© 2011 Glumm et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielfältigt, verbreitet und öffentlich zugänglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Gliederung

Text

Objective: Over the last year we have established a new co-culture method combining murine motor cortical and spinal slices. Until now, no comparable trial has been described. We have intensely investigated motor cortical regeneration and sprouting to the spinal cord and characterized the ingrowing neurons and invading cells. Currently, we are analyzing the influence of potentially enhancing compounds on neuronal reestablishment in our experimental setup.

Methods: The spinal cord (sc) was dissected from non-fluorescent C57Bl/6 mice postnatal day 0 until 3 (P0-P3). The motor cortex (mc) was extracted from Bl6.beta-actin green fluorescent protein (GFP) pups (P0-P3). The medial zone of the mc was orientated on a semipermeable cell culture, inserted into the rostral end of the sc and co-cultured for up to 14 days. To analyze the effects of potential outgrowth compounds, different concentrations of Coomassie Brilliant Blue G-250 (BBG), cyclosporin A (CSA) and beta-nerve growth factor (NGF) were added to the incubation medium directly after preparation (Peng et al., 2009; Sofroniew et al., 2001; Bonnici et al., 2009). Using different immunhistochemical stainings after tissue fixation, we characterized all ingrowing GFP cells in the spinal cord as well as counted and compared ingrowing GFP fibers in the treatment groups. To evaluate neuronal recovery a cross-sectional stack profile of every slice was recorded.

Results: Our data show ingrowing fibers and growth cones that are already detectable after 1 day in vitro (DIV). Measuring the speed of fiber growth via confocal microscopy, we could verify a decrease of ingrowth in the course of the incubation time. In addition, immunhistochemical stainings after 3 and 6 DIV suggest a strong neuronal outgrowth, a reestablishment of cortical fibers and new connections. Additionally, we detected cortical microglia and astrocytes migrating into spinal tissue during incubation. Furthermore, we confirmed the beneficial effects of CSA on cortical regeneration and could exclude BBG and NGF from enhancing neuronal recovery in our experimental setup.

Conclusions: CSA induced outgrowth and reestablishment of motor cortical neurons in our spinal cord – motor cortex co-culture model. Thus, signal transduction is involved in motor cortical recovery. Our in vitro method offers possibilities to test axon-regenerative properties of determined compounds as well as other treatments and provides an important tool to answer a variety of questions in the field of neuronal regeneration.