不同基底硬度在调控HepaRG细胞分化为肝细胞样细胞中的作用  

Role of different substrate elasticity in regulating rapid differentiation of HepaRG cells into Hepatocyte-like cells

作　　者：李甲志 王冰洁 劳建乐 徐静[1] 

机构地区：[1]广西医科大学第一附属医院肝胆外科,南宁市530021 [2]清华大学生命科学院,清华大学医学院生物医学工程系,北京市100084

出　　处：《实用医学杂志》2017年第11期1739-1743,共5页The Journal of Practical Medicine

基　　金：国家自然科学基金资助项目(编号:81360372)

摘　　要：目的:探讨基底硬度在调控肝祖细胞系Hepa RG细胞分化为肝细胞样细胞的可能性,为生物人工肝提供肝细胞来源。方法:实验分为4组,分别采用白蛋白-绿色荧光蛋白-报告系统、Image J软件检测细胞ALB的表达;倒置显微镜观察细胞形态;Alamar blue检测细胞数量。结果:在第4小时,4s组细胞的ALB分别与8s组、16s组、Glass组比较,差异无统计学意义(t=0.791,1.389,2.481,P>0.05);在第4天,4s组分别与16s组和Glass组比较,差异均有统计学意义(t=12.410,12.520,P<0.05),4s组与8s组比较,差异没有统计学意义(t=2.603,P>0.05);在第7天,4s组分别和8s组、16s组、Glass组比较,差异均有统计学意义(t=3.266,6.725,8.005,P<0.05)。显微镜观察结果显示Hepa RG细胞在分化前呈单一肝祖细胞样形态,分化后呈现肝细胞样和胆管细胞样形态。Alamar blue检测结果显示在第4和7天,4s组的细胞总数分别与8s组、16s组、Glass组比较,差异均无统计学意义(P>0.05)。结论:软的基底更有利于HepaRG细胞分化。Objective To investigate the potential of substrate elasticity in regulating rapid differentiation of HepaRG cells into hepatocyte-like cells, and further provide hepatocytes for bioartificial liver. Methods The substrate elasticity was divided into 4 groups. The expressions of albumin （ALB） were detected by albumin-green fluorescent protein-reporter system （ALB-GFP-reporter system） and Image J software; the cell morphology was observed by microscope and the amounts of cell were detected by cell Titer-Blue cell viability assay kit （alamar blue）. Results The results of ALB showed that at the 4th hour, the expressions of ALB inside the HepaRG cells between 4s group and 8s group, 16 s group and Glass group were not statistically different （t = 0.791,1.389, 2.481, P 〉 0.05） ; at the 4th day, the expressions of 4s group had statistical differences in comparison with those of 16s group and Glass group （t = 12.41, 12.52, P 〈 0.05） , but not statistical difference when compared with those of 8s group （t = 2.603, P 〉 0.05） ; at the 7th day, the expressions of 4s group were statistically different from those of 8s group, 16s group and Glass group （t = 3.266, 6.725, 8.005, P 〈 0.05）. The microscope showed that only a typical epithelial morphology was observed before differentiation, then hepatocyte-like cells and bile duct-like cell were observed at differentiated phases. The alamar blue showed that at the 4th and the 7th day, the cell number of 4s group were not statistically different from that of 8s group, 16s group and Glass group （P 〉 0.05）. Conclusion Soft substrate can promote differentiation of HepaRG cells.

关 键 词：生物人工肝 HepaRG细胞 肝细胞样细胞 基底硬度 

分 类 号：R575[医药卫生—消化系统]