氧化三甲胺在细胞超低温冻存中的应用
首发时间:2017-02-24
摘要:以天然渗透压调节剂氧化三甲胺(TMAO)作为超低温冻存保护剂,采用超速冷冻方法对两种模型细胞-人肺癌细胞(GLC-82)以及人宫颈癌细胞(Hela)进行超低温保存。差示扫描量热(DSC)测试表明,TMAO可以显著降低水溶液的冰点,抑制冰晶生成,有利于保护细胞免受冰晶损伤;细胞毒性测试结果表明,细胞在添加1% TMAO的细胞培养液中浸泡3天后,未观察到细胞受到毒性损伤;细胞冻存测试表明,当TMAO浓度为6%(w/v,TMAO/培养基)时,冻存效果最优,GLC-82细胞与Hela细胞的复苏成活率分别达到64%和41%。
关键词: 细胞生物学 细胞超低温保存 冻存保护剂 氧化三甲胺 超速冷冻保存 差示扫描量热技术
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The application of trimethylamine oxide (TMAO) in cell cryopreservation
Abstract:The natural osmolyte trimethylamine oxide (TMAO) was tested as a novel cryoprotectant to cryopreserve two types of model cells (GLC-82 cells and Hela cells) with an ultra-rapid freezing protocol. The differential scanning calorimetry (DSC) tests revealed that TMAO had a strong ability to decrease the freezing point of aqueous solution, indicating its strong inhibition to water crystallization, which could protect cells from ice injury. The cytotoxicity assay suggested that TMAO was nontoxic when we exposed GLC-82 cells to 1% TMAO solution for three days. Furthermore, at the optimal concentration of TMAO, the post-thaw viability of GLC-82 cells and Hela cells could reach 64% and 41%, respectively. In conclusion, TMAO could potentially be a nontoxic and high efficient cryoprotectant for cell storage, transportation, as well as clinical cell therapy.
Keywords: cell biology cell cryopreservation cryoprotectant trimethylamine oxide ultra-rapid freezing differential scanning calorimetry
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