Department of Medical Simulation Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.
Cell Transplant. 2010;19(6):691-9. doi: 10.3727/096368910X508780. Epub 2010 Jun 3.
Cryoprotective agents (CPAs) such as dimethyl sulfoxide (DMSO), glycerol, ethylene glycol, and propylene glycol have been used for the cryopreservation of cells and tissues. DMSO is the most effective CPA but shows high cytotoxicity and can effect differentiation. ɛ-poly-L-lysine (PLL) derivatives show higher cryopreservation efficiency than the conventional CPAs. Culture medium solutions with 7.5 w/w% of PLL whose amino groups of more than 50 mol% were converted to carboxyl groups by succinic anhydride showed higher postthaw survival efficiency of L929 cells than those of current CPAs without the addition of any proteins. In addition, rat mesenchymal stem cells were cryopreserved more effectively than with DMSO and fully retained the potential for proliferation and differentiation. Furthermore, many kinds of cells could be cryopreserved with PLL having the appropriate ratio of COOH groups, regardless of the cell types, including adhesive and floating cells, human- and mouse-derived cells, primary cells, and established cell lines. The properties might be associated with the antifreeze protein properties. These results indicate that these polymeric extracellular CPAs may replace current CPAs and the high viability after thawing and nonnecessity of serum ensure that these CPAs may be used in various preservation fields.
冷冻保护剂（CPAs），如二甲基亚砜（DMSO）、甘油、乙二醇和丙二醇，已被用于细胞和组织的冷冻保存。DMSO 是最有效的 CPA，但具有高细胞毒性，并可能影响分化。ɛ-聚-L-赖氨酸（PLL）衍生物比传统的 CPA 具有更高的冷冻保存效率。由琥珀酸酐将其氨基中超过 50 mol%的 PLL 转化为羧基的浓度为 7.5 w/w%的培养基溶液显示出比不添加任何蛋白质的当前 CPA 更高的 L929 细胞冻后存活率。此外，大鼠间充质干细胞的冷冻保存效果比 DMSO 更好，并且完全保留了增殖和分化的潜力。此外，许多种细胞可以用 PLL 进行冷冻保存，PLL 的 COOH 基团比例合适，无论细胞类型如何，包括贴壁细胞和悬浮细胞、人源和鼠源细胞、原代细胞和已建立的细胞系。这些特性可能与抗冻蛋白的特性有关。这些结果表明，这些聚合物细胞外 CPA 可能取代现有的 CPA，并且解冻后的高存活率和对血清的非必要性确保了这些 CPA 可用于各种保存领域。
