Abstract: In order to effectively reduce the concentration of dimethyl sulfoxide (Me2SO, DMSO), reduce its toxicity as a cryoprotectant for cryopreservation of cells, and improve cell survival rate, this experiment targeted hepatocytes and adopts response surface method for cryoprotection. The formulation was optimized based on response surface methodology. The results showed that the optimal ratio of cryoprotectant was DMSO concentration 3%, glycerol concentration 6%, trehalose concentration 0.1%. Under this condition, hepatocyte rewarming survival rate was 84.35%, which could effectively reduce the concentration of DMSO. The result reduced the toxic damage that caused to the liver cells and laid the foundation for subsequent research.

Key words: hepatocyte, toxicity, DMSO, response surface method, survival

摘要： 为有效降低干细胞冷冻保护液中的二甲基亚砜(Me2SO，DMSO)浓度，减少其作为冷冻保护剂对细胞低温保存时产生的毒性，提高细胞存活率，以肝细胞为目标，采取响应面法对冷冻保护剂配方进行了优化。结果表明：冷冻保护剂的最佳配比为DMSO浓度3%、甘油浓度6%、海藻糖浓度0.1%，在此条件下肝细胞复温存活率可达到84.35%，研究结果有效地降低了DMSO的浓度，减少了由此带来的对肝细胞造成的毒性损害，为后续的研究奠定了基础。

关键词: 肝细胞, 毒性, 二甲基亚砜, 响应面法, 存活率