Isolation And Characterization Of Mesenchymal Stem Cells From Adult Human Liver

Objective:The aim of the study was to isolate and characterize mesenchymal stem cells from adult human liver(aLHMSCs).We also investigate the effects of aLHMSCs on the activated hepatic stellate cells.Methods:We isolated aLHMSCs by Dispase-collagenase four-step perfusion method,density gradient centrifugation and adherence screening method.Utilizing flow cytometry,immunofluorescence and RT-PCR,the phenotype and genes of aLHMSCs were analyzed.Furthermore we investigated the capacity of aLHMSCs to differentiate into adipogenic,osteogenic and hepatocyte-like cells.The effects of aLHMSCs on HSC-Li(an immortalized human hepatic stellate cell type)were also explored by indirect co-culture.Results:We successfully isolated aLHMSCs,which were fibroblast-like and expressed mesenchymal stem cell markers(CD73,CD29,CD105,CD44 and CD90).However,They did not express hematopoietic stem cell markers(CD34,CD45,CD117 and CD 13 3),hepatic progenitor cell markers(CD 19,AFP,CD326,CD117 and CD34)and hepatic stellate cell markers(CK19 and HLA-? class molecule),suggesting that they were not hematopoietic origin and absolutely different from hepatic progenitor cells and hepatic stellate cells.They were also positive for a stem cell marker(vimentin)and hepatocyte-specific markers(CK18,CK8 and ALB),indicating that they owned properties of stem cells and hepatic cells.In addition,aLHMSCs were able to differentiate into adipogenic,osteogenic and hepatic cells.After hepatic induction,hepatocyte-specific genes were up regulated and functions were strengthened(such as glycogen storage,low-density lipoprotein uptake,CYP450,albumin synthesis and urea secretion).Moreover,aLHMSCs could inhibit the activation of HSC-Li through secreting cytokines,an effect TNF-a involved in.Conclusion:Adult human liver is a novel origin of mesenchymal stem cells.Cells isolated were of general characterizations of mesenchymal stem cells and able to generate functional hepatocyte-like cells.Moreover,aLHMSCs inhibited the activation of hepatic stellate cells through secreting cytokines.Thus,aLHMSCs may be a new alternative for cell therapy of liver diseases.