【摘要】 目的:观察匀速与变速旋转培养对微胶囊HepLL永生化人肝细胞、HepG2肝细胞聚集体形成和活性的影响。方法:一步法海藻酸钠—壳聚糖(AC)微胶囊包裹HepG2及HepLL细胞,随机分为两大组,每组按转速分3小组。动态观察并测量各组聚集体大小、数目变化,ELISA动态检测各组白蛋白合成量,比色法检测氨的清除量,高效液相色谱法检测安定转化功能。结果:与匀速旋转培养组相比,变速旋转培养组微胶囊细胞第6、8、10、12、14、16天聚集体平均个数、平均最大径、白蛋白合成量、氨清除量以及安定转化量均明显提高(均P<0.01),微胶囊HepLL各组在各个时间点的白蛋白合成量、氨清除量及安定转化功能均优于HepG2(均P<0.01)。结论:微胶囊转瓶变速培养显著加快微胶囊HepLL、HepG2细胞聚集体形成并提高细胞活性,未来HepLL有可能代替HepG2用于人工肝治疗。更多还原

【Abstract】 Objective:To observe the effect of uniform and shift rotation culture on the formation and activity of the alginate-chitosan(AC) microencapsulated HepLL immortalized human hepatocytes and HepG2 cells aggregates.Methods:AC microcapsulated HepG2 and HepLL cells were randomly divided into two groups.Each group was divided into 3 subgroups according to uniform and shift rotation culture.The size and number of aggregates were observed and measured under laser confocal microscopy and inverted microscope dynamically.The amount of albumin synthesis was detected by ELISA,the clearance of ammonia was detected by colorimetry,and diazepam conversion function was detected by high performance liquid chromatography(HPLC).Results:On day 6,8,10,12,14 and 16,the number and size of the aggregates,albumin synthesis,diazepam clearance and ammonium clearance increased significandy in shift rotation culture group than in uniform group(all P<0.01).The albumin synthesis,diazepam clearance,and ammonium clearance in the microencapsulated HepLL groups were significantly higher than those of HepG2 cells at any time(all P <0.01).Conclusion:Shift rotation culture can significantly promote the formation and increase the activity of AC microencapsulated HepLL and HepG2 aggregates,and HepLL cells may be more suitable for bioartificial liver than HepG2.更多还原