Day, AGE;
Bhangra, KS;
Murray-Dunning, C;
Stevanato, L;
Phillips, J;
(2017)
The Effect of Hypothermic and Cryogenic Preservation on Engineered Neural Tissue.
Tissue Engineering Part C: Methods
, 23
(10)
pp. 575-582.
10.1089/ten.TEC.2017.0244.
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Abstract
This study explored different approaches to preserve Engineered Neural Tissue (EngNT), a stabilised cellular collagen hydrogel containing columns of aligned Schwann cells for nervous system repair. The ability to preserve EngNT without disrupting cellular and extracellular components and structures is important for clinical translation and commercialisation. Stabilised cellular gels and EngNT constructs were preserved under various conditions and cell survival assessed using live/dead microscopy and metabolic assay. Optimal survival was recorded in hypothermic (4ºC) conditions for 2-3 days using Hibernate®-A media, and, for longer term cryogenic storage (liquid nitrogen), using a mixture of 60% Dulbecco's Modified Eagle's Medium media (DMEM), 30% foetal calf serum (FCS) and 10% dimethyl sulfoxide (DMSO). Functionality and structure of preserved EngNT was assessed in co-culture with dorsal root ganglion (DRG) neurons, which indicated that alignment of Schwann cells and the ability of EngNT to support and guide neuronal regeneration were not disrupted. The identification of conditions that preserve EngNT will inform development of storage and transport methodologies to support clinical and commercial translation of this technology and other therapies based on cellular hydrogels.
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