重庆医学2017，Vol.46Issue(17)：2305-2308,4.DOI:10.3969/j.issn.1671-8348.2017.17.001
利用海藻酸水凝胶构建仿肝板肝组织三维共培养模型
Construction of 3D co-culture model of liver tissue mimicking hepatic plate constructed by alginate hydrogel
摘要
Abstract
Objective To construct a new 3D co-culture model of hepatic cells.Methods The alginate hydrogel microfiber was prepared with sodium alginate,microfluidic chip,liver cell line C3A and human umbilical vein endothelial cell line EA.hy926.The experimental group was the mimick hepatic plate group (F),meanwhile the mixed disorderly hydrogel microfiber was prepared as the control group (H).The live cell double fluorescent labeling was used to verify the cell arrangement structure in the microfiber.Microfibers were cultured for 1 week,the microfiber morphologies were daily observed and the hepatocyte activity was measured,the albumin (Alb),alanine aminotransferase (ALT),aspartate aminotransferase (AST),lactate dehydrogenase (LDH-L),alpha 1 antitrypsin (α1AT),coagulation factor (F Ⅶ),glutathione S transferase alpha 1 (GSTα1) and cytochrome P450 1A2 (CYP1A2) contents in the cultures were detected.Results In the group,C3A cells in hydrogel arranged in two or three lines at the center,EA.hy926 cells located in the bilateral sides of C3A cells,showing the hepatic plate arrangement;in the group H,the two kinds of cells were mixed together in a state of disorder.The liver tissue cords were formed on 3 d.The hydrogel microfiber diameter of two groups showed little changes over time without statistical significance(P＞0.05).The viability of hepatocytes reached a maximum on 5 d in the group F and on 6 d in group H,while the group F was higher than group H at each time point except on 1 d.The albumin secretion in the two groups showed basically same trend,and reached a maximum on 3 d,then began to decline on 4 d.The levels of ALT,AST and LDH-L in the group F were decreased to a minimum on 3 d,and then varied trend after 4 d was the same as the group H.The α1AT at each time point except on 5 d had statistically significant difference between the two groups(P＜ 0.05);the GSTα1 secretion amount was increased over time in both groups,which at each time point in the group F was significant ly higher than that in the control group (P＜0.05).The FⅦ secretion amount in the group F was gradually increased before 7 d,while which in the control group continued to decline from 2 d,and which in the group F was significantly higher than that in the control group H from 3 d(P＜0.05).The change of intracellular CYP1A2 content with time lapse was unobvious in the group H,while which from 4 d in the group F began to be significantly higher than that in the group H(P＜0.05).Conclusion The three dimensional co-culture model of liver tissue mimicking hepatic plate is successfully constructed,in which the hepatocyte function is expected to be maintained for a long time.关键词
海藻酸钠/水凝胶/微流体/肝细胞/肝板/三维共培养Key words
alginate/hydrogel/microfluidic/hepatocyte/hepatic plate/3D co-culture分类
医药卫生引用本文复制引用
王高尚,贾志栋,李阳,彭青,高毅..利用海藻酸水凝胶构建仿肝板肝组织三维共培养模型[J].重庆医学,2017,46(17):2305-2308,4.基金项目
国家高技术研究发展计划(863计划)资助(2012AA020505) （863计划）
国家自然科学基金资助项目(81470875) （81470875）
广东省省级科技计划项目(2014B020227002) （2014B020227002）
广东省省级科技计划项目(2015B090903069) （2015B090903069）
广东省自然科学基金资助项目(2014A030312013) （2014A030312013）
广东省省级科技计划项目(2015B020229002) （2015B020229002）
广东省省级科技计划项目(2013B091100001) （2013B091100001）
南方医科大学开发培育项目(CL033042). （CL033042）
