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BACKGROUND:Previous studies have confirmed that bone marrow mesenchymal stem cells in vitro can promote hepatic stel ate cellapoptosis and inhibit its activity, in which the mechanism of action remains unknown. OBJECTIVE:To screen out apoptosis-related genes during hepatic stel ate cellapoptosis regulated by bone marrow mesenchymal stem cells using gene chip technology. METHODS:Purified human bone marrow mesenchymal stem cells were seeded in 6-wel Transwel plate and cocultured with hepatic stel ate cells. Cultured human bone marrow mesenchymal stem cells alone served as control group, and cultured for 72 hours. The alterations in apoptosis-related genes were analyzed between culture alone group and coculture group using gene chip technology. The genes strongly associated with regulation of hepatic stel ate cells were selected. RESULTS AND CONCLUSION:By the functional classification of second-generation SABiosciences Gene chips, apoptotic gene screening found that after coculture, significantly upregulated genes in bone marrow mesenchymal stem cells contained:AKT1, PIK3R2, DAPK1, DHCR24, NOTCH2 and BDNF. Combined with previous findings, we hypothesized that NOTCH may play a key role in the regulation of hepatic stel ate cells by bone marrow mesenchymal stem cells.
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Objective To analyze the short-term and long-term effectiveness of radiofrequency ablation combined with transcatheter arterial chemoembolization (TACE) for the treatment of hepatocellular carcinoma (HCC).Methods The clinical data of 70 HCC patients who had received thermal ablation (group A) done or in combination with TACE (group B) were retrospectively analyzed.Results The rate of intrahepatic distant recurrence in group B (25 cases) was lower than that in group A (45 cases) (X2 =3.845,P =0.046) and the tumor-free survival rate was higher than group A (X2 =5.020,P =O.030).There were no differences in the local tumor progression rate (X2 =0.853,P =0.374) and overall survival (x2 =2.316,P =0.154) between two groups.Incidence of bone marrow suppression in group B was higher than that of group A (X2 =5.642,P =0.042).Major complications didn't occur in any group(X2 =2.016,P =0.183).The costs was higher(t =7.738,P ＜0.001) and the hospital stay was longer (t =5.921,P =0.003) in group B than group A.Conclusions Compared with ablation alone,combined therapy is able to reduce short-term recurrence,and improve tumor-free survival.Combine therapy is safe and effective method for small liver carcinoma.
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BACKGROUND:It is reported that bone marrow mesenchymal stem cell(BMSC)transplantation might be a promising treatment for liver fibrosis.But the mechanism is still unclear.OBJECTIVE:To observe the hepatic stellate cells apoptosis induced by BMSC transplantation,and to study the mechanism of BMSC in treating hepatic fibrosis in vivo.METHODS:CCl_4 subcutaneous injection was performed to induce rat liver fibrosis.After 8 weeks of CCU injection,20 rats which underwent successful model establishment were randomly divided into experimental group and control group,10 in each group.The experimental group received MSC transplantation via tail vein injection,and the control group were given DMEM instead.The rats were killed and the livers were harvested at three time point,the day of MSC transplantation,3 days after transplantation,and 7 days after transplantation.The hydroxyproline content was detected by HE and Masson staining,and the expression changes of α-smooth muscle actin(α-SMA)proteins were determined using immunohistochemistry.The apoptosis of hepatic stellate cells were determined by α-SMA and TUNEL(terminal dUTP nick-end labeling)dual-staining.RESULTS AND CONCLUSION:After 8 weeks of CCU injection,the hydroxyproline content increased and histology indicated progress of liver fibrosis.At 7 days after MSC transplantation,the hydroxyproline in the liver was decreased,and the liver fibrosis was alleviated in the experimental group but aggravated in the control group.Immunohistochemistry indicated that α-SMA positive cells were increased at 8 weeks after CCU injection.At day 7 after transplantation,α-SMA positive cells in the experimental group were significantly less than control group(P ＜ 0.05).At 3 days after transplantation,the hepatic stellate cells apoptosis in the experimental group was significantly aggravated compared with control group(P ＜ 0.05).This suggested that MSC transplant was an effective treatment for liver fibrosis.MSC inducing hepatic stellate cells apoptosis may be one of the mechanisms.
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Objective To construct the bioartificial liver by bone mesenchymal stem cell (BMSC) and alginate scaffold. Method Alginate scaffold was used as the cell carrier for the cultivation of BMSC and the differentiation from BMSC into hepatic like cells was induced by the cell factors of HGF, EGF and FGF-4 in the scaffold in vitro. The compatibility of the cells and the scaffold was observed by microscopy and the function of the differentiatd cells was tested. The gene of AFP and ALB was detected by RT-PCR. The secretion of ALB and the urea synthesis of the cells were tested by ALB kit and urea kit respectively. The glycogen synthesis and the CK-18 was tested by the glycogen stanning method and the immunofluorescence test. Results BMSC was able to attach, grow and proliferate well in the alginate scaffold, the well compatibility was observed by microscopy. ALB and urea were detected in the cultivating medium, the gene of ALB and AFP was identified by RT-PCR. The glycogen synthesis ability and the expression of CK-18 were induced during the differentiation. Conclusion The three dimensional atginate scaffold exhibited well compatibility with BMSC, BMSC could be differentiated into the hepatic like cell in the scaffold. BMSC and the alginate scaffold could be used to construct the bioartificial liver for the hepatic tissue engineering.
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BACKGROUND: There is no accepted treatment for liver fibrosis recently. Bone marrow meaenchymal stern cells (BMSCs) used in the treatment of liver fibrosis has been reported as an effectively treatment, but the mechanism is unclear.OBJECTIVE: To study the regulation of hepatic stellate cells mediated by human BMSCs in vitro.DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the Center for Stem Cells and Tissue Engineering of Sun Yat-sen University and the Central Laboratory of Third Affiliated Hospital of Sun Yat-sen University from June to December 2008.MATERIALS: Human bone marrow masenchymal stem cells were collected from normal youth volunteers; Human hepatic stellate cells and normal liver call line L-O2 were supplied by the Animal Experimental Center of Sun Yat-sen University.METHODS: The purified human BMSCs and hepatic stellate calls were set up in Transwell co-culture system. The incubation density was 2×104cells/well. L-O2 was set up instead of human BMSCs as negative control. Hepatic stellate cells cultured alone served as blank control group. The culture was performed for 72 hours.MAIN OUTCOME MEASURES: Morphology of hepatic stellate cells and results of immunocytochemical staining. Apoptosis of hepatic stellte calls was determined by flow cytometry. Western blot were used to assay the expression of α-actin.RESULTS: Activated hepatic stellate cells presented fiat and thin shape under an inverted microscope. Fat drop was lack in cytoplasm, a -actin located in hepatic stellate calls, with the presence of high tension fibers. Compared with the L-O2 + hepatic stellate cell and hepatic stellate call groups, the apoptotic rate of hepatic stellate cells was significantly increased in the BMSC + hepatic stellate cell group (P < 0.05). α -actin expression was significantly down-regulated.CONCLUSION: Human BMSCs can inhibit activation of hepatic stellate ceils and promote them apoptosis, which may be the anti-hepatic fibrosis mechanism of BMSCs.
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especially in the group of ceasing CCl4 injection. Conclusion After transplanted into the liver by spleen injection, hepatic oval cells could plant in the portal area, which improve liver function and alleviate the cirrhosis level.
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Objective To compare the effect of hepatic oval cells and bone marrow mesenchymal stem cells to treat liver fibrosis in mice. Methods The C57BL/6 mice were fed with 3,5-diethoxycarbonyl-1, 4-dihydro-collidine (DDC) in a standard food at a concentra-tion of 0.1% to activate the oval cells proliferation, then the nonparenchymal cells were separated by in situ perfusion and density gradient centrifugation, and finally Sca-1 antibody was used in conjunction with magnetic activated cell sorting (MACS) to separate the Sca-1 +cells. Other mice were injected subcutaneously with CCl4 to establish experimental liver fibrosis model. Finally, the fibrosis mice were trans-planted with HOC or MSC or saline by spleen injection according to the group they belonged to. Four weeks later, the liver function index, the hydroxyproline and the histopathological change among the groups were used to evaluate the effect of the treatment. Result The liver function index was improved after transplantation. HOC and MSE treatment could reduce both the hydroxyproline level and alleviate the fibro-sis, but HOC seems more effective in the group with continuing CCl4 injection(P < 0.05). When stopping injecting CCl4, the change of fi-brosis was not visible(P >0.05). Conclusion Both hepatic oval cells and bone marrow mesenchymal stem cells can improve the liver function, and alleviate the fibrosis level by transpleen transplantation, but hepatic oval cells were more effective.