The Effect of Hypothermic and Cryogenic Preservation on Engineered Neural Tissue.
The Effect of Hypothermic and Cryogenic Preservation on Engineered Neural Tissue.
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DOI：
10.1089/ten.tec.2017.0244
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发表时间：
2017-10
期刊：
影响因子：
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通讯作者：
Phillips JB
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文献类型：
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作者：
Day AGE;Bhangra KS;Murray-Dunning C;Stevanato L;Phillips JB
这项研究探索了保存工程神经组织 (EngNT) 的不同方法，这是一种稳定的细胞胶原水凝胶，含有排列整齐的雪旺细胞柱，用于神经系统修复。在不破坏细胞和细胞外成分和结构的情况下保留 EngNT 的能力对于临床转化和商业化非常重要。稳定的细胞凝胶和 EngNT 构建体在各种条件下保存，并使用活/死显微镜和代谢测定评估细胞存活率。使用 Hibernate®-A 培养基在低温 (4°C) 条件下记录 2-3 天的最佳存活率，对于长期低温储存（液氮），使用 60% Dulbecco 改良 Eagle 培养基、30% 胎牛血清和 10% 二甲亚砜的混合物。在与背根神经节神经元共培养中评估了保留的 EngNT 的功能和结构，这表明雪旺细胞的排列以及 EngNT 支持和引导神经元再生的能力没有被破坏。确定保存 EngNT 的条件将为存储和运输方法的开发提供信息，以支持该技术和基于细胞水凝胶的其他疗法的临床和商业转化。
This study explored different approaches to preserve engineered neural tissue (EngNT), a stabilized, 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 commercialization. Stabilized 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 (4°C) 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, 30% fetal bovine serum, and 10% dimethyl sulfoxide. Functionality and structure of preserved EngNT were assessed in coculture with dorsal root ganglion 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.