The Effect On Hepatocellular Function In 3D Co-culture Of Liver Tissue Mimicking Hepatic Plate

Background:Liver failure is caused by a variety of factors(such as viruses,alcohol,drugs,etc.),resulting in a large number of liver cells necrosis,leading to severe liver dysfunction or decompensation,and a group of clinical syndrome such as coagulation disorders,jaundice,liver encephalopathy,ascites are the main signs.At present,liver transplantation is an effective method for the treatment of liver failure,but the world’s organ transplantation is facing a serious shortage of sources.To this end,artificial liver technology and liver tissue engineering targetting on the function repair,replacement,reconstruction of the liver in liver failure treatment provides a new choice.Objective:We combined with liver tissue engineering and bioartificial liver technology to simulate the basic structure of normal liver,and proposed a study on the three-dimensional(3D)co-culture technology in liver tissue mimicking hepatic plate.The hepatocyte function was measured in this culture method,and compared with other in vitro culture methods,in order to improve the function of hepatocyte in vitro and to be suitable for the bioartificial liver.In addition,this three-dimensional culture of liver tissue is also expected to become an effective evaluation tool for drug development and testing.Methods:By using microfluidic,hydrogel,soft lithography and other techniques,humanized liver cell and endothelial cell co-culture,we successfully constructed an imitation of liver tissue structure.To study the effect on hepatocellular function in 3D co-culture of liver tissue mimicking hepatic plate,we designed in vitro functional experiments and animal therapy experiments.Compared with mixed three-dimensional co-culture,sandwich co-culture,common plate co-culture methods,morphology,viability and function(albumin,alanine aminotransferase,coagulation factor,etc.)of liver cell were observed and tested in two weeks of the experiment.The male Kunming mice were treated with D-galactosamine and lipopolysaccharide in acute liver failure model.The mice were treated with intraperitoneal injection of liver tissue mimicking hepatic plate.To comprehensively evaluate the treatment effect of the liver tissue on the mice,the survival rate and the blood biochemical indexes of Kunming mice were observed and tested.Results:There was no significant difference in the morphology between the hepatic plate group and the mixed three-dimensional group.The hepatocyte viability in the hepatic plate group was higher than that in the mixed three-dimensional group.The CYP1A2,coagulation Factor Ⅱ,testosterone metabolism,diazepam metabolism in the hepatic plate group were higher than the other three groups.In the animal experiment,the treatment effect of the hepatic plate group was obvious,compared with the control group,and the survival rate of the mice was significantly improved,the majority of biochemical indicators decreased significantly.Conclusions:The effect on hepatocellular function in 3D co-culture of liver tissue mimicking hepatic plate was manifested in all aspects.Compared with sandwich co-culture and common plate co-culture methods,liver cell metabolism,protein synthesis and secretion,drug metabolism,etc.in the hepatic plate group were much better,which is worth further study.Liver tissue in the alginate hydrogel could replace partially the liver function of Kunming mice,and also had some improvement in the blood biochemical indexes.In this study,humanized cells were used to provide some reference value for bioartificial liver,drug development and testing and cell transplantation.