苦参碱对于人肝细胞冻存保护效应的实验性研究

目的:了解苦参碱对人肝细胞液氮冻存复苏后功能的影响,探索一种改善人肝细胞冻存的新方法。方法:将胎肝细胞及常规培养的L-02人肝细胞系分别以5种不同冻存保护体系犤10%二甲亚砜(DMSO)、5%DMSO+0.2mg/L苦参碱、5%DMSO+2mg/L苦参碱、5%DMSO+6mg/L苦参碱、5%DMSO+10mg/L苦参碱犦在液氮中冷冻保存1个月。1个月后快速复苏,进行存活率、形态学及功能检测。结果:不同冻存保护体系保存的人肝细胞存活率、形态学及功能表现均有所不同。其中5%DMSO+6mg/L苦参碱组效果最好,复苏后细胞存活率、24h贴壁率及细胞功能检测优于其他冻存液组。结论:⑴苦参碱对人肝细胞液氮冻存有保护作用,6mg/L苦参碱是最好的浓度;⑵苦参碱与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。

Protective effect of matrine on human hepatocytes preserved in liquid nitrogen

Objective To study the effect of matrine on the function of the revived human hepatocytes preserved in liquid nitrogen and to expore a better method for the cryopreservation of hepatocytes. Methods Human fetal hepatocytes and immortalized L- 02 hepatocytes were stored in liquid nitrogen for one month with five different cryoprotectants (10% DMSO alone, 5% DMSO 0.2 mg/L matrine, 5% DMSO 2 mg/L matrine, 5% DMSO 6 mg/L matrine and 5% DMSO 10 mg/L matrine). After one month, the cells were thawed rapidly and their livability, morphology and function were measured. Results The human hepatocytes preserved in different cryoprotectants showed different livability, morphological manifestation and cell fuction. Among the five methods, a combination of 5% DMSO 6 mg/L matrine was the best. The livability, 24- hour adherent rate and function of the cells cryopreserved with 5% DMSO 6 mg/L matrine were better than those of the cells with other different cryoprotectants (P< 0.05). Conclusions First, matrine has a protective effect on the human hepatocytes preserved in liquid nitrogen and reaches its highest level at a concentration of 6 mg/L. Second, matrine also has a synergistic effect on DMSO, and the use of matrine combined with DMSO can reduce the concentration of DMSO.