Cell & Tissue Systems, Inc., North Charleston, SC 29406, USA.
Cryobiology. 2012 Jun;64(3):240-4. doi: 10.1016/j.cryobiol.2012.02.006. Epub 2012 Feb 15.
Dimethylsulfoxide, the most commonly employed cryoprotectant for cells, has well documented cytotoxic effects in patients. Among the compounds available that may provide protection to cells and tissues during preservation with less cytotoxicity is trehalose. Some animals, such as brine shrimp and tardigrades, accumulate trehalose during periods of extreme environmental stress. In this study, experiments were performed to evaluate the effects of culturing a bovine endothelial cell line (ATCC #CCL-209) in the presence of trehalose prior to preservation by freezing. A number of factors were shown to contribute to cell retention of metabolic activity and proliferative potential including cell culture time with trehalose and the solution conditions during cryopreservation. Using an optimized protocol consisting of 24 h of cell culture with 0.2 M trehalose followed by cryopreservation with 0.2-0.4 M trehalose in sodium bicarbonate buffered Eagles minimum essential medium at pH 7.4 resulted in 87±4% post-preservation cell metabolic activity expressed as relative fluorescence based upon reduction of resazurin to resorufin. This new method provides an alternative preservation strategy to the more classical preservation methods employing dimethylsulfoxide available for cells and tissues.
二甲基亚砜是最常用的细胞冷冻保护剂，已被充分证明对患者具有细胞毒性作用。在可用的化合物中，海藻糖可能在保存过程中提供对细胞和组织的保护，同时具有较低的细胞毒性。一些动物，如盐水虾和缓步动物，在极端环境压力期间积累海藻糖。在这项研究中，进行了实验以评估在冷冻保存之前用海藻糖培养牛内皮细胞系（ATCC #CCL-209）对其的影响。已经证明许多因素有助于细胞保持代谢活性和增殖潜力，包括用海藻糖进行细胞培养的时间和冷冻保存过程中的溶液条件。使用由 24 小时用 0.2 M 海藻糖进行细胞培养和用 0.2-0.4 M 海藻糖在 pH 值为 7.4 的碳酸氢钠缓冲的 Eagles 最小必需培养基中进行冷冻保存组成的优化方案，导致保存后细胞代谢活性保持在 87±4%，表示为基于还原 Resazurin 为 Resorufin 的相对荧光。这种新方法为细胞和组织提供了替代更经典的二甲基亚砜保存方法的保存策略。
