Phillips, J. B.
A tissue engineered 3D culture model for studying damage and repair in peripheral nerves.
In: 2011 PNS Biennial Meeting of the Peripheral Nerve Society, 25-29 June 2011, Potomac, Maryland, US.
An advanced cell culture system has been developed that can be used to examine interactions between neurons and glia and explore their responses to damage, overcoming some of the limitations associated with conventional culture systems and animal models. Here we report the methodology for generating tissue engineered nerves in vitro and demonstrate the potential of the technology for use in PNS research. Schwann cells can be seeded within a collagen solution that sets to form a gel within a rectangular mould. By tethering the gel at opposing ends, the contraction mediated by the Schwann cell interaction with the collagen is restrained in the longitudinal direction, causing the cells to align parallel to the axis of tension. Neurons seeded within this environment extend neurites from one end of the gel to the other, guided and supported by the anisotropic glial environment. The resulting construct therefore comprises aligned neurites growing in contact with aligned Schwann cells within a collagen environment. The Schwann cells can myelinate the neurites and the subsequent model system mimics many of the key components of the endoneurium of a peripheral nerve. The cell culture environment is highly controllable and can be adjusted to simulate various damage and disease states. It is also amenable to a wide range of analyses including fluorescence, confocal, time lapse and electron microscopy, protein and RNA measurement and electrophysiological recording. Outputs that can be quantified include neurite length and alignment, Schwann cell viability and alignment, myelination, deposition of extracellular matrix molecules and release of cytokines. Our group has used cultured dorsal root ganglion neurons from wild type and transgenic rats in this system to develop devices for the repair of peripheral nerve trauma, for screening the ability of potentially therapeutic glial cells to support and myelinate regenerating axons, and to examine the effect of targeted cancer therapies on peripheral nerves. Having developed robust protocols for generating reproducible model nerves we are currently exploring how to use this medium-throughput approach more widely to simulate peripheral neuropathy and facilitate the development of therapeutic interventions.
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