Reparation Of Mouse Actue Liver Injury By Bone Marrow Mesenchymal Stem Cells

Objective To explore whether human bone marrow derived mesenchymal stem cells (MSCs) are able to differentiate into hepatocyte-like cells under appropriate condition in vitro. To induce mouse acute liver injury model by intraperitoneal injection with 10% CCl₄ in mouse and evaluate the therapeutic efficacy by injecting BMSCs and explore the possible mechanisms.Methods The human BMSCs were isolated and cultured. The BMSCs were induced by hepatocyte growth factor (HGF), basic fibroblast growth factor (bFGF) and co-cultured with damaged mouse liver tissues respectively. The specific genes of hepatocyte were detected by RT-PCR and real-time PCR in different culture period. The hepatic markers were identified by immunofluorescent staining. Mouse acute liver injury model was induced by intraperitoneal injection with 10% carbon tetrachloride (CCl₄) in mouse. After injection of BMSCs into the livers, RT-PCR for the human 17a gene was used to locate exogenous hMSCs in mouse livers, and the specific genes of hepatocyte were detected. Peripheral blood and liver specimens were collected at 7, 14, and 21 days after transplantation. The serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) was measured, and the pathological state of the hepatocytes was assessed, to investigate the therapeutic efficacy of MSCs as well as the possible mechanisms about it.Results Expression of alpha-fetoprotein (AFP), cytokeratin-18, (CK-18) and tryptophan 2,3-dioxygenase (TDO) were not detectable until day 7 and increased with time. Differentiated cells, which were induced by HGF and bFGF at 7 days, were positive for the hepatocyte-specific markers, alpha-fetoprotein (AFP), albumin (ALB), hepatocyte nuclear factor (HNF), human serum albumin (HSA), and tryptophan 2,3-dioxygenase (TDO). After being co-cultured with damaged liver tissues of mouse separated by a transwell membrane, the shape of MSCs turned round, AFP and TDO were detected in cells for 21 days. Mouse acute liver injury model was established successfully. The human 17a satellite DNA sequence was amplified in mouse heart, kidney, lung, brain, spleen and different parts of mouse liver in the experiment for fetal MSCs location. The levels of serum ALT and AST decreased significantly inexperimental groups than in control groups (P<0.05). The liver recoverysituation in experimental groups was better than in control groups. AFP and TDO were detected in mouse liver of experimental groups by RT-PCR.Conclusions Human bone marrow MSCs are able to differentiate into hepatocyte-like cells under culture with HGF,bFGF and injured liver tissues. Exogenous MSCs could home to the injured liver and cure mouse liver CCl4-injury efficiently by differentiating. These investigations could offer original experimental model and evidence about efficient cure of MSCs to liver disorders such as acute injury , serve a new source of cells for cell therapy of hepatic diseases, and also open up a new path to them.