Expression Of NF-?B And Ubc9 On Inducing Differentiation Of Bone Mesenchymal Stem Cell To Hepatocyte

Objective In order to research BMSCs induced and differentiated into hepatocytes suitable conditions and inducing differentiation mechanism of NF-?B and Ubc9 in vitro and in vivo,and identified the hepatocytes to provide an experimental groundwork for the therapy of hepatic diseases using BMSCs transplantion.Methods BMSCs were isolated from bone marrow of rats,then cultured and expanded in DMEM F12 with 10%NCS.We provided BMSCs the hepatic microenvironment using some factors from mouse liver such as G(0.5g/L)and CM,and compared with the different culture factors such as HGF(30?g/L),?-NGF(50?g/L),and RA(0.3g/L)by immunocytochemistry,negative control group without inducer.The cell morphology was observed after induced 14d,21d under a microscope at any time,?l-antitrypsin(AAT)protein of liver cell-specific detected and expression of NF-?B and cell Ubc9 protein was observed by immunofluorescence staining.Results(1)Microscopic observation showed that part of cells after 48h visible fusiform,irregularly shaped,after significantly increased the number of adherent cells,fusiform cells,polygonal,uneven distribution of the colony was like distribution trends,more uniform distribution of cells after passage,consistent cell morphology after P3,fusiform or spindle-shaped,was swirling like arrangement or parallel(2)BMSCs induced morphological observation process BMSCs showed endothelial cells in most shuttle type or kind of fibroblast-like elongated,tightly adherent than the average 9-12 d bottom covered with boards or culture bottle.A variety of ingredients added to induce the cells after induction 7d fusiform morphology significantly,a small amount of cells showed round,inducing 14d,21 d after the spindle or fibroblast-like cells induced with the extension of time,become spindle,irregular round or similar liver-like cells.(3)Immunocytochemistry results Showed that Compared with the control group,the expression of ?l-antitrypsin protease(AAT)are positive inducing by G,CM,HGF,P-NGF RA after 14d,21d,which HGF,P-NGF,G and CM group stronger than RA group(P<0.05).AAT-positive cells in the form of becoming shorter round gradually.Immunofluorescence results also showed that compared with the control group,the expression of NF-?B-induced liver cell a gradual upward trend with the change process,inducing by CM,G,?-NGF,HGF and RA after 14d,21d,Ubc9 protein expression mainly in the cytoplasm.Compared to the group,after 14d,21d the expression of NF-?B and Ubc9 proteins,and inducing by G,CM,HGF,?-NGF group are stronger stronger than RA group.Conclusion:BMSCs can differentiate into hepatocytes inducing by G,CM ?-NGF,HGF and RA.As long as the offer or cytokine microenvironment can promote the differentiation of BMSCs to hepatocytes.NF-?B and Ubc9 were positive expression induced differentiation process,which studies the molecular biological mechanisms of bone marrow mesenchymal stem cells induced to differentiate into hepatic cells provide new understanding of.