Castro Vânia I B, Craveiro Rita, Silva Joana M, Reis Rui L, Paiva Alexandre, C Duarte Ana Rita
3B's Research Group - Biomaterials, Biodegradable and Biomimetic, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Avepark, 4805-017, Barco, Guimarães, Portugal; ICVS/3B's PT Government Associated Laboratory, 4710-243, Braga/Guimarães, Portugal.
LAQV@REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal.
Cryobiology. 2018 Aug;83:15-26. doi: 10.1016/j.cryobiol.2018.06.010. Epub 2018 Jun 23.
Natural deep eutectic systems (NADES) are mostly composed of natural primary metabolites such as sugars, sugar alcohols, organic acids, amino acids and amines. These simple molecules have been identified in animals living in environments with extreme temperature amplitudes, being responsible for their survival at negative temperatures during winter. Herein, we report for the first time the use of NADES based on trehalose (Treh) and glycerol (Gly) in cryopreservation, as cryoprotective agents (CPA). The evaluation of the thermal behaviour of these eutectic systems, showed that NADES have a strong effect on the water crystallization/freezing and melting process, being able to reduce the number of ice crystals and hence ice crystal damage in cells, which is a crucial parameter for their survival, upon freezing. Using this NADES as CPA, it is possible to achieve similar or even better cellular performance when compared with the gold standard for cryopreservation dimethyl sulfoxide (DMSO). In this sense, this work relates the physical properties of the NADES with their biological performance in cryopreservation. Our comprehensive strategy results in the demonstration of NADES as a promising nontoxic green alternative to the conventional CPA's used in cryopreservation methods.
天然低共熔体系（NADES）大多由天然初级代谢产物组成，如糖类、糖醇类、有机酸、氨基酸和胺类。这些简单分子已在生活于温度振幅极大环境中的动物体内被发现，它们在冬季低温环境下对动物的存活起着重要作用。在此，我们首次报道了基于海藻糖（Treh）和甘油（Gly）的NADES在冷冻保存中作为冷冻保护剂（CPA）的应用。对这些低共熔体系热行为的评估表明，NADES对水的结晶/冻结和融化过程有显著影响，能够减少冰晶数量，从而减少细胞在冷冻时因冰晶造成的损伤，这是细胞存活的关键参数。将这种NADES用作CPA时，与冷冻保存的金标准二甲基亚砜（DMSO）相比，能够实现相似甚至更好的细胞性能。从这个意义上讲，这项工作将NADES的物理性质与其在冷冻保存中的生物学性能联系起来。我们的综合策略证明了NADES是冷冻保存方法中传统CPA的一种有前景的无毒绿色替代品。
