Experimental Studies On Treatment Of Liver Injury In HBV Mice By Inducing And Differentiating Bone Marrow Mesenchymal Stem Cells Into Hepatocyte Cells

Background and Aims：Liver disease is one of the most common diseases that affect human health.Liver injury have been caused by various harmful factors：viral hepatitis, alcoholic hepatitis, anddrug-induced hepatitis, the clinical manifestations have liver necrosis, fatty liver, cholestasis, liverfibrosis, cirrhosis, and liver cancer. The current treatment of liver injury is still a grim global issues，but the stem cell transplantation provided a better treatment for this kind of diseases. Bone marrowmesenchymal stem cells(BMSCs) is a kind of cells which have multiple differentitationpotentialitiesand strong self-replication ability.It is expected to be a new seed cells which can be usedto treat liver injury.we induced BMSCs to hepatocyte-like cells by HGF and EGF, transplantedhepatocyte cells into the model of liver injury, determined the effect of cell transplantation throughthe identification.The research results can provide experimental basis for clinical application.Method:(1) The establishment and comparative study of HBV transgenic mice and C57BL/6mice bonemarrow mesenchymal stem cells(BMSCs) isolated and cultured system respectively.(2) Treated purified BMSCs by using HGF and EGF，then using immunocytochemical staining todetected the expression of liver cell surface markers，such as CK18(cytokeratin18) and PAS(3) We made the model of liver injury, transplanted the BMSCs which marked into model,using thefluorescence microscope to observe the colonization situation of the BMSCs,and examined theliver function.Results：(1) C57BL/6mice primary BMSCs grow rapidly, then it was swirling growth according to the1:3passage, which of morphology similar to fibroblasts. HBV transgenic mice primary BMSCs grewslowly, according to1:2passage, which of growth is slow, and have a strong dependence ofserum and cell density.(2) Treated purified BMSCs by using HGF and EGF，then using immunocytochemical staining todetected the expression of liver cell surface markers，such as CK18(cytokeratin18) and PAS，theresults are postive.(3) In serum levels of ALT, AST， the model group A2and the control group A1were significantlydifference (P <0.05); at the end of the4th weeks of the model group A2and experimental groupB1、B2were significantly difference（P<0.05）；between experimental group B1and B2weresignificantly difference at the end of the4th weeks（P<0.05）Conclusion：(1) Compared with C57BL/6mice, HBV transgenic mice BMSCs existed in serum-dependent cellgrowth and density dependence in vitro.(2) BMSCs induced to hepatocyte cells by HGF and EGF.(3) From the level of serum of ALT,AST and the results of fluorescent markers，between the groupof BMSCs and BMSCs induced to hepatocyte cells were significantly difference at the end ofthe4th weeks，hepatocyte cells group was better than others to treat liver injury.