Qingdao University, Qingdao, 266071, China; Department of Cardiovascular Surgery, The Affiliated Hospital of Qingdao University, Qingdao, 266000, China.
Department of Cardiovascular Surgery, The Affiliated Hospital of Qingdao University, Qingdao, 266000, China.
Cryobiology. 2019 Feb;86:103-110. doi: 10.1016/j.cryobiol.2018.11.002. Epub 2018 Nov 17.
Cryopreservation of tissue cells is an important method to maintain cell viability and cellular function. However, cell viability and function are less than ideal by conventional cell cryopreservation methods, which may result in apoptosis and necrosis of cells in cryopreservation. Trehalose plays a role in maintaining cell structure and protecting cells from stress. However, owing to the difficulty in transport of trehalose across the cell membrane, its antifreeze effect is limited. A large amount of trehalose (up to 237 ± 8.5 mM) can be delivered to smooth muscle cells incubated in a medium containing trehalose and apatite nanomaterials at 37 °C for 6 h. Our data showed that trehalose was efficiently delivered intracellularly with the aid of nanoparticles (NP), with a loading efficiency up to 137.3 ± 34.5%, thus allowing for cryopreservation of LMC with nontoxic sugar as the sole cryoprotectant. Colloidal bioelastic apatite NP were used as bioactive promoters for the cryopreservation of tissue cells with trehalose. The addition of apatite NP in the medium substantially increased aortic smooth muscle cell cryosurvival, up to 83.6% (30% improvement over control without NP), a level comparable to that associated with the traditional MeSO cryoprotective regimen. Furthermore, the cytotoxicity of nanocapsules in the intracellular delivery of trehalose was negligible. This method provides a new option to enhance the activity of valvular cells for cryopreservation.
组织细胞的低温保存是维持细胞活力和细胞功能的重要方法。然而，传统的细胞低温保存方法使细胞活力和功能不太理想，可能导致细胞在低温保存过程中发生凋亡和坏死。海藻糖在维持细胞结构和保护细胞免受应激方面发挥作用。然而，由于海藻糖难以穿过细胞膜运输，其抗冻效果有限。在含有海藻糖和磷灰石纳米材料的培养基中，将大量海藻糖（高达 237±8.5mM）在 37°C 下孵育 6 小时，可以输送到平滑肌细胞中。我们的数据表明，纳米颗粒（NP）可有效地将海藻糖递送到细胞内，装载效率高达 137.3±34.5%，从而可以使用无毒糖作为唯一的冷冻保护剂来冷冻保存 LMC。胶体生物弹性磷灰石 NP 被用作含有海藻糖的组织细胞低温保存的生物活性促进剂。在培养基中添加磷灰石 NP 可大大提高主动脉平滑肌细胞的冷冻存活率，达到 83.6%（比没有 NP 的对照组提高 30%），与传统的 MeSO 冷冻保护方案相当。此外，纳米胶囊在细胞内递送海藻糖的细胞毒性可以忽略不计。该方法为增强瓣膜细胞的冷冻保存活性提供了一种新的选择。
