O'Rourke, Caitriona; Loughlin, Jane; Drake, Rosemary and Phillips, James
Optimising glial cell alignment in 3D culture to facilitate the development of neural tissue models for CNS research.
In: Tissue Engineering and Regenerative Medicine International Society - World Congress 2012, 5 - 8 September 2012, Austria, Vienna.
Aim: The overall aim is to develop robust models for neuroscience research, with neural cells arranged in a hydrogel matrix to resemble living nervous system tissue. We create engineered neural tissue by a process of initial glial cell self-alignment within tethered 3D collagen gels and the aligned glia then support and direct neuronal growth to recreate the anisotropy of an organised CNS tract.
Method: A two-stage approach was developed to determine the glial cell seeding density to achieve consistent predictable alignment regardless of the cell source. (1) Contraction profiles were established using C6 glioma cells at densities from 0.1 to 6 million cells/ml in freefloating round collagen gels, in simple 24-well and 96-well plate assays. (2) Chosen seeding densities were then used to assess the degree of cellular alignment using tethered rectangular collagen gels.
Results: By combining data from the contraction profiles and alignment assays, the relationship between the % contraction of the free-floating round gels and the extent of cellular alignment in the tethered rectangular gels was established.
Conclusion: We have shown that contraction profiles in simple multiwell plate assays, using a small number of cells, can efficiently assess glial cells from different sources and determine the optimal seeding density for generating robust anisotropic engineered neural tissue.
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