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O'Rourke, Caitriona; Drake, Rosemary; Loughlin, Jane and Phillips, James
(2013).
URL: http://firmsymposium.com/files/4213/8497/1590/Cait...
Abstract
Aim: The overall aim is to develop robust culture models that recreate the 3D environment of the CNS, allowing neurons and glial cells in vitro to behave similarly to their counterparts in vivo. This approach is being used to develop a simple, consistent model system which can be used at a scale suitable for drug screening, providing an experimental platform for neurobiological research.
Methods: Engineered neural tissue is created by a process of initial glial cell self-alignment within a tethered 3D collagen hydrogel. The aligned glia can then support and direct neuronal growth to recreate the anisotropic architecture of an organised CNS tract. RAFT technology (www.raft3dcellculture.com) is employed to stabilise the cellular hydrogels, through partial removal of interstitial fluid thereby increasing matrix and cell density. Initial experiments characterised the cell and matrix parameters required for reliable, consistent cellular alignment in large (1ml) moulds using C6 glioma cells, then scaled down moulds were assessed in terms of cellular alignment and support of neurite growth.
Results: Miniaturised moulds can be used to generate anisotropic cellular collagen gels, which can be stabilised using RAFT. Glial cell alignment in our scaled down system displayed similar patterns as in the larger standard system, with over 50% angles of deviation in both middle and side regions being less than 30°.
Discussion: The results indicate that engineered neural tissue models can be scaled down without compromising glial cell alignment and stabilisation. This approach may be beneficial for the development of an experimental platform for widespread adoption as it minimises the cell numbers required for generating robust anisotropic engineered neural tissue and could also lead to an increase in throughput to match the requirements for drug discovery screening.
CORE (COnnecting REpositories)
CORE (COnnecting REpositories)
