School of Materials Science and Engineering, Tianjin Key Laboratory of Composite and Functional Materials, Tianjin University, Tianjin 300350, China.
Tianjin Blood Center, Tianjin 300110, China.
J Mater Chem B. 2022 Jun 15;10(23):4452-4462. doi: 10.1039/d2tb00445c.
Cryopreservation of human erythrocytes suitable cryoprotectants is essential for transfusion during emergencies, but the conventional glycerolization method requires a tedious thawing-deglycerolization process. Alternatively, trehalose, a nonreducing disaccharide, has gained much attention as a biocompatible cryoprotectant due to its nature in living organisms capable of surviving extreme cold and desiccation. In this work, cryopreservation of human erythrocytes was realized through high intracellular trehalose enhanced by benzyl alcohol at 4 °C with membrane stabilization of maltotriose-grafted ε-poly(L-lysine). Intracellular trehalose could reach 94.2 ± 12.1 mM with slight impacts on morphology and cell functions, and the post-storage cryosurvival of human erythrocytes could achieve 96.2 ± 3.4% membrane protection by the glycopeptide. It has been demonstrated that the functional glycopeptide performed as an extracellular cryoprotectant accompanied by high intracellular trehalose for synergistic cryopreservation of human erythrocytes in the biocompatible glycerol-free conditions. This two-step approach involving augmentation of intracellular trehalose at a hypothermic temperature and membrane stabilization of the functional glycopeptide could be an alternative way for human cell cryopreservation.
人类红细胞的低温保存 合适的冷冻保护剂对于紧急情况下的输血至关重要，但传统的甘油化方法需要繁琐的解冻-去甘油化过程。另一方面，海藻糖作为一种非还原二糖，由于其在能够耐受极端寒冷和干燥的生物体内的性质，作为一种生物相容性的冷冻保护剂而受到广泛关注。在这项工作中，通过在 4°C 下用苯甲醇增强细胞内海藻糖，并通过麦芽糖三糖接枝的 ε-聚（L-赖氨酸）稳定细胞膜，实现了人类红细胞的低温保存。细胞内海藻糖可达到 94.2±12.1mM，对形态和细胞功能的影响很小，糖肽可实现 96.2±3.4%的红细胞低温保存后存活率，从而实现细胞膜的保护。研究表明，该功能糖肽作为一种细胞外冷冻保护剂，与细胞内高浓度的海藻糖协同作用，可在无甘油的生物相容性条件下对人类红细胞进行协同低温保存。这种两步法涉及在低温下增加细胞内海藻糖的含量，并稳定功能糖肽的细胞膜，这可能是人类细胞低温保存的一种替代方法。
