Monitoring glial cell behaviour and neuron-glial interactions in 3D hydrogel and aggregate culture systems

Phillips, J. and Loughlin, J. (2011). Monitoring glial cell behaviour and neuron-glial interactions in 3D hydrogel and aggregate culture systems. In: 10th European Meeting on Glial Cells in Health and Disease, 13-17 Sep 2011, Prague, Czech Republic.




We use a variety of three-dimensional culture systems to study normal glial cell function and neuronal-glial interactions as well as modelling pathological conditions in vitro. Reactive gliosis is difficult to model in two dimensions since astrocytes in monolayer systems tend to exhibit a reactive phenotype continuously. We have shown that astrocytes in 3D collagen hydrogels adopt a less reactive phenotype than in equivalent monolayer cultures. On stimulation using TGF-beta1, the astrocytes become ramified and hypertrophic and express characteristic reactivity markers, mimicking the astrocyte response to damage in vitro. Coupled with confocal microscopy and image analysis, this system provides a versatile test-bed to monitor astrocyte responses in simulated damage and repair scenarios. Three-dimensional cocultures of neurons and glia have permitted the effect of astrocyte alignment on neurite regeneration to be studied, and have allowed us to simulate the inhibitory interfaces that form at sites of CNS damage or the PNS/CNS boundary.
The process of CNS myelinogenesis, which involves complex physical and biochemical interactions between neurones and cells of the oligodendroglial lineage at different stages of maturation, is another example of a process that is poorly modelled in two dimensions. We have used rotation-mediated reaggregate cell cultures and 3D collagen hydrogel cultures to study aspects of this dynamic process in vitro, in particular analysing changes in the protein composition of oligodendroglial lipid rafts during myelinogenesis. Our culture models also allow us to study demyelination in vitro, using inflammatory cytokines or antimyelin antibodies as triggers.
Our studies confirm the value of 3D cell culture systems as an important complementary approach to both traditional monolayer culture systems and in vivo models; permitting cell–cell and cell–matrix interactions that approximate more closely to those in vivo than is observed in the former, whilst enabling a level of observation and manipulation that is impossible in the latter.

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