Chemically Induced Extracellular Ice Nucleation Reduces Intracellular Ice Formation Enabling 2D and 3D Cellular Cryopreservation.
Chemically Induced Extracellular Ice Nucleation Reduces Intracellular Ice Formation Enabling 2D and 3D Cellular Cryopreservation.
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DOI：
10.1021/jacsau.3c00056
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发表时间：
2023-05-22
期刊：
影响因子：
8
通讯作者：
Whale, Thomas F
中科院分区：
文献类型：
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作者：
Murray, Kathryn A;Gao, Yanan;Griffiths, Christopher A;Kinney, Nina L H;Guo, Qiongyu;Gibson, Matthew I;Whale, Thomas F
3D 细胞组装体（例如球体）比传统 2D 单层细胞更准确地再现体内状态，并且正在成为减少或取代动物测试的工具。目前的冷冻保存方法并未针对复杂的细胞模型进行优化，因此它们不容易存储，也不像二维模型那样广泛使用。在这里，我们使用可溶性冰成核多糖使细胞外冰成核并显着改善球体冷冻保存结果。这比单独使用 DMSO 更能保护细胞，其主要优点是成核剂在细胞外发挥作用，因此不需要渗透到 3D 细胞模型中。悬浮液、2D 和 3D 冷冻保存结果的严格比较表明，高温冰成核减少了（致命的）细胞内冰的形成，并且在 2/3D 模型的情况下，这减少了相邻细胞之间冰的传播。这表明细胞外化学成核剂可以彻底改变先进细胞模型的存储和部署。
3D cell assemblies such as spheroids reproduce the in vivo state more accurately than traditional 2D cell monolayers and are emerging as tools to reduce or replace animal testing. Current cryopreservation methods are not optimized for complex cell models, hence they are not easily banked and not as widely used as 2D models. Here we use soluble ice nucleating polysaccharides to nucleate extracellular ice and dramatically improve spheroid cryopreservation outcomes. This protects the cells beyond using DMSO alone, and with the major advantage that the nucleators function extracellularly and hence do not need to permeate the 3D cell models. Critical comparison of suspension, 2D and 3D cryopreservation outcomes demonstrated that warm-temperature ice nucleation reduces the formation of (fatal) intracellular ice, and in the case of 2/3D models this reduces propagation of ice between adjacent cells. This demonstrates that extracellular chemical nucleators could revolutionize the banking and deployment of advanced cell models.