Polyampholytes as Cryoprotective Agents for Mammalian Cell Cryopreservation
Polyampholytes as Cryoprotective Agents for Mammalian Cell Cryopreservation
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DOI：
10.3727/096368910x508780
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发表时间：
2010-01-01
影响因子：
3.3
通讯作者：
Hyon, Suong Hyu
中科院分区：
文献类型：
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作者：
Matsumura, Kazuaki;Bae, Jung Yoon;Hyon, Suong Hyu
冷冻保护剂（CPA）如二甲基亚砜（DMSO）、甘油、乙二醇和丙二醇已用于细胞和组织的冷冻保存。DMSO是最有效的CPA，但显示出高细胞毒性并可影响分化。ε-聚-L-赖氨酸（PLL）衍生物显示出比常规CPA更高的低温保存效率。与7.5 w/w%的PLL，其氨基基团的超过50摩尔%的琥珀酸酐转化为羧基基团的培养基溶液显示出更高的解冻后的存活效率的L929细胞比那些目前的CPA没有添加任何蛋白质。此外，大鼠骨髓间充质干细胞的冻存效果优于DMSO，并充分保留了增殖和分化的潜力。此外，许多种类的细胞可以用具有适当比率的COOH基团的PLL冷冻保存，而与细胞类型无关，包括粘附和漂浮细胞、人和小鼠来源的细胞、原代细胞和已建立的细胞系。这些性质可能与抗冻蛋白的性质有关。这些结果表明，这些聚合物细胞外CPA可以取代现有的CPA，解冻后的高活力和不需要血清，确保这些CPA可以用于各种保存领域。
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. epsilon-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.