Construction of 3D co-culture model of liver tissue mimicking hepatic plate constructed by alginate hydrogel
Wang Gaoshang;Jia Zhidong;Li Yang;Peng Qing;Gao Yi;Second Department of Hepatobiliary Surgery,Zhujiang Hospital,Southern Medical University/State Key Laboratory of Organ Failure Research/Co-Innovation Center for Organ Failure Research/Guangdong Provincial Research Center of Artificial Ogan and Tissue Engineering;
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 C3 Aand 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 1week,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 1antitrypsin(α1AT),coagulation factor(FⅦ),glutathione S transferase alpha 1(GSTα1)and cytochrome P450 1A2(CYP1A2)contents in the cultures were detected.Results In the group,C3 Acells in hydrogel arranged in two or three lines at the center,EA.hy926 cells located in the bilateral sides of C3 Acells,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 3d.The hydrogel microfiber diameter of two groups showed little changes over time without statistical significance(P0.05).The viability of hepatocytes reached a maximum on 5d in the group F and on 6din group H,while the group F was higher than group H at each time point except on 1d.The albumin secretion in the two groups showed basically same trend,and reached a maximum on 3d,then began to decline on 4d .The levels of ALT,AST and LDH-L in the group F were decreased to a minimum on 3d,and then varied trend after 4d was the same as the group H.Theα1AT at each time point except on 5d had statistically significant difference between the two groups(P0.05);the GSTα1secretion amount was increased over time in both groups,which at each time point in the group F was significantly higher than that in the control group(P0.05).The FⅦ secretion amount in the group F was gradually increased before 7d,while which in the control group continued to decline from 2d,and which in the group F was significantly higher than that in the control group H from 3d(P0.05).The change of intracellular CYP1A2 content with time lapse was unobvious in the group H,while which from 4d in the group F began to be significantly higher than that in the group H(P0.05).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.
【Fund】： 国家高技术研究发展计划(863计划)资助(2012AA020505);; 国家自然科学基金资助项目(81470875);; 广东省省级科技计划项目(2014B020227002);广东省省级科技计划项目(2015B090903069);广东省省级科技计划项目(2015B020229002);广东省省级科技计划项目(2013B091100001);; 广东省自然科学基金资助项目(2014A030312013);; 南方医科大学开发培育项目(C1033042)
【CateGory Index】： R318.08;R657.3
【CateGory Index】： R318.08;R657.3