Spinal cord organotypic slice cultures for the study of regenerating motor axon interactions with 3D scaffolds

Jose Gerardo-Nava*, Dorothee Hodde, Istvan Katona, Ahmet Bozkurt, Torsten Grehl, Harry W. M. Steinbusch, Joachim Weis, Gary A. Brook

*Corresponding author for this work

    Research output: Contribution to journalArticleAcademicpeer-review


    Numerous in-vitro techniques exist for investigating the influence of 3D substrate topography on sensory axon growth. However, simple and cost-effective methods for studying post-natal motor axon interactions with such substrates are lacking. Here, spinal cord organotypic slice cultures (OSC) from postnatal day 7-9 rat pups were presented with spinal nerve roots, or blocks of fibrin hydrogel or 3D microporous collagen scaffolds to investigate motor axon substrate interactions. By 7-14 days, axons from motor neuronal pools extended into the explanted nerve roots, growing along Schwann cell processes and demonstrating a full range of axon-Schwann cell interactions, from simple ensheathment to concentric wrapping by Schwann cell processes and the formation of compact myelin within a basal lamina sheath. Extensive motor axon regeneration and all stages of axon-Schwann interactions were also supported within the longitudinally orientated microporous framework of the 3D collagen scaffold. In stark contrast, the simple fibrin hydrogel only supported axon growth and cell migration over its surface. The relative ease of demonstrating such motor axon regeneration through the microporous 3D framework by immunofluorescence, two-photon microscopy and transmission electron microscopy strongly supports the adoption of this technique for assaying the influence of substrate topography and functionalization in regenerative bioengineering.
    Original languageEnglish
    Pages (from-to)4288-4296
    Issue number14
    Publication statusPublished - May 2014


    • Nerve regeneration
    • Collagen
    • Fibrin
    • In vitro test
    • Scaffold

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