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Natural deep eutectic systems for nature‐inspired cryopreservation of cells
AIChE Journal ( IF 3.7 ) Pub Date : 2020-10-02 , DOI: 10.1002/aic.17085
Kathlyn Hornberger 1 , Rui Li 1 , Ana Rita C. Duarte 2 , Allison Hubel 3
Affiliation  

Natural deep eutectic systems (NADES) are emerging as potential cryoprotective agents (CPA) for cell preservation. In this investigation, we develop an optimized CPA formulation using trehalose‐glycerol NADES (T:G) diluted in Normosol‐R and supplemented with isoleucine. Differential scanning calorimetry (DSC) is used to define the thermophysical properties of NADES‐based solutions, and Raman spectroscopy is used to characterize the effect of NADES on ice formation and hydrogen bonding. Jurkat cells are cryopreserved in each solution, and post‐thaw cell recovery, apoptosis, and growth are quantified. Raman spectra and heat maps show that NADES suppresses both ice formation and dehydration of the nonfrozen region. Supplementing NADES with isoleucine does not affect the solution's thermophysical properties but significantly improves the cells' survival and proliferation post‐thaw. The study indicates that thermophysical properties of CPA solutions alone cannot predict optimal cell survival, suggesting that stabilization of biological structures by CPAs may play a role in successful cryopreservation.

中文翻译:

天然深共晶系统,用于自然冷冻细胞

天然深层共晶系统(NADES)逐渐成为细胞保存的潜在防冻剂(CPA)。在这项研究中,我们使用在Normosol-R中稀释并补充有异亮氨酸的海藻糖-甘油NADES(T:G)开发了一种优化的CPA配方。差示扫描量热法(DSC)用于定义基于NADES的溶液的热物理性质,拉曼光谱用于表征NADES对冰形成和氢键的影响。将Jurkat细胞冷冻保存在每种溶液中,并对融化后的细胞恢复,凋亡和生长进行定量。拉曼光谱和热图表明,NADES抑制了非冻结区域的冰块形成和脱水。用异亮氨酸补充NADES不会影响溶液的热物理性质,但会显着改善细胞的 解冻后的存活和增殖。该研究表明,仅CPA溶液的热物理性质无法预测最佳的细胞存活,这表明CPA稳定生物结构可能在成功的低温保存中起作用。
更新日期:2020-10-02
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