糖醇类物质对人肝细胞低温保存的影响

Cryopreservation of Human Hepatocyte by Sugar-alcohol based CPAs

将传统肝细胞冻存保护剂配方中的DMSO的浓度减为5%,并添加不同浓度的D-山梨醇、木糖醇或麦芽糖醇,置于-80℃的冰箱冻存两周,两周后将细胞快速复温,进行细胞存活率、24h贴壁率以及细胞形态学的检测,并与10%浓度DMSO组对照比较。结果表明,D-山梨醇、木糖醇、麦芽糖醇均对人肝细胞的低温保存有一定的保护作用,其中5%DMSO+0.4M D-山梨醇组复温后细胞的存活率、贴壁率均优于其他冻存液组,D-山梨醇与DMSO联合使用降低了冻存保护剂DMSO的浓度,表明两者有协同作用。

Different types of sugar alcohol is usually used as cryoprotective agents （CPAs） to preserve human hepatocytes. In this study, a new types of sugar alcohol based CPAs with low toxicity was explored by replacing Me2SO with D-sorbic alcohol, xylosic alcohol and maltol. The Me2SO concentration in CPAs was reduced to 5%. Human hepatocytes attached to plastic cover slips were treated with the above CPAs and then placed in the fridge at -80℃ for two weeks. The survival rate, 24h cell detachment and cell morphology were tested and compared with the results using 10% Me2SO as CPAs. The results show that 5% Me2SO with D-sorbic alcohol gives the best results. It is concluded that the new CPAs are effective to human hepatocytes cryopreservation.