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[Abstract] Objective To investigate the effect of wingless-type MMTV integration site family member 5b(Wnt5b) gene overexpression mediated by recombinant adenovirus on the differentiation of mouse embryonic liver stem cells and repair of chronic liver injury in mice. Methods Recombinant adenoviruses expressing Wnt5b and green fluorescent protein (GFP) were applied respectively to infect mouse fetal liver stem cells HP14-19, and induced its differentiation and verified the expression of Wnt5b through Real-time PCR and Western blotting. It also applied indocyanine grean(ICG) uptake experiment and periodic acid-schiff(PAS) staining to detect the differentiation ability of HP14-19 into hepatocyte-like cells. The Real-time PCR was chosen to detect hepatocyte markers albumin (Alb) and cytokeratin 18 (CK18) expression. Forty-eight experimental male BALB/ c mice were randomly divided into control group, model group, stem cell treatment group and Wnt5b modified stem cell treatment group. The carbon tetrachloride(CCl
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Objective: To investigate the effects of hepatic stem cells survival in the pathological microenvironment of cholestatic cirrhosis, and the effects of sodium glycosaminodeoxycholate (GCDC) on apoptosis of hepatic stem cells in vitro. Methods: Balb/c mice were subjected to bile duct ligation (BDL) to simulate the pathological environment of cholestatic cirrhosis; Liver stem cells HP 14-19 were transplanted back into liver by the splenic vein and survival of cell colonization was detected; Effects of sodium glycosaminodeoxycholate on the viability of HP 14-19 cells at different concentrations by cell counting kit-8(CCK-8) and crystal violet staining, flow cytometry was used to detect the apoptosis of HP14-19 cells treated with 600 μmol/L GCDC 24 hours later, and the expression of Bax, Bcl-2, Capase-3, phosphorylated adenosine 5′-monophosphate-activated protein kinase α (p-AMPKα) and adenosine 5′-monophosphate-activated protein kinase α (AMPKα) were detected by Western blotting. Results: The results of CCK-8 and crystal violet staining showed that the proliferation of HP 14-19 cells was inhibited with the increase of the concentration of sodium glycosaminodeoxycholate. Flow cytometry showed that the apoptosis rate of GCDC treated group was higher than that of control group. Compared with the control group, the expression of Bax, Capase-3 was up-regulated and the expression of Bcl-2 protein was down-regulated in the experimental group. The results showed that GCDC could induce apoptosis of HP14-19 (P<0.05) AMPK was activated. Conclusion: Microenvironment of cholestatic cirrhosis induced apoptosis of liver stem cells.
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Objective To analyze the clinical and pathological characteristics of 35 hepatocellular carcinoma (HCC) patients with bile duct tumor thrombi (BDTT),and to investigate the expressions of CD133,CD90,EpCAM,CK19,VEGF,and C-kit in the tumor tissues.Methods 35 HCC patients with BDTT out of 943 HCC patients who underwent surgical treatment were studied.The expressions of biomarkers in tissue specimens were determined by immunohistochemistry.35 HCC patients without BDTT were selected using the method of stratified sampling as a control group.Results In 19 of 35 patients,the diameters of the primary tumor were less than 5 cm (range 0 ～ 17 cm,average 6.9 ± 0.7 cm).When compared to the control group,most of the primary tumors were moderately to lowly differentiated (33/35,94％ vs 18/ 35,51％),had incomplete capsules (18/35,51％ vs 3/35,8％) and micro vascular invasion (29/35,83％ vs 7/35,20％).The positive expression rates of CD90,EpCAM,CK19,VEGF,CD133,and C-kit in the group of patients with HCC with BDTT and in the control group were 82.9％,77.1％,71.4％,85.7％,80.0％,80.0％ and 57.1％,54.3％,34.3％,65.7％,54.3％,51.4％,respectively.The 1-,2-,3-year postoperative survival rates of the HCC patients with BDTT were 69％,37％,20％ respectively which were worse than the HCC patients without BDTT (1-,2-,3-year postoperative survival rates were 88％,72％,62％ respectively,P ＜ 0.05).Conclusions The prognosis of HCC patients with BDTT was worse than HCC patients without BDTT.The expressions of liver stem cell biomarkers in the tumor specimens of the group of HCC patients with BDTT were higher than the control group.These findings prompt that this kind of HCC originate from liver stem ceils.
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Objective To investigate the effect of hepatopoietin Cn(HPPCn) on liver stem cells.Methods In this study, WB-F344 cell line was used, and MTT and flow cytometry assay were conducted to determine cell proliferation and apoptosis.Transwell assay was used to test the migration of WB-F344 cells.A 2AAF-partial hepatectomy(PH) mouse model was used to observe the effect of HPPCn on liver stem cell proliferation in vivo.Results HPPCn enhanced WB-F344 cell proliferation and migration and activated the SphK1, Erk and Stat3 signal pathways.The analysis of the 2AAF-PH mouse model showed that oval cells in the experimental group far outnumbered those in control and the regeneration of the liver was improved post PH.Conclusion HPPCn can increase the liver stem cell proliferation and survival while promoting the regenenation of the liver by augmenting oval cell proliferation.
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ObjectiveTo provide decision-making information support for recognizing research trends, selecting frontier science and technology, shaping reasonable science and technology distribution, a visualization analysis was applied to the international liver stem cell research. Methods Based on bibliometric method and visualization analysis tools, various fields including publication years, countries of publications, journals and organizations were analyzed exhaustively on publications of international liver stem cells which were searched in Science Citation Index Expanded (SCIE).In addition, details on main subjects, evolution and research fronts were presented in this paper. Results 1092 publications were searched in SCIE,which were growing rapidly since 2000.The production of China is far behind United States of America and Japan. Publications were distributed in concentrated journals but involved multidisciplinary. Liver stem cells research evolution basically experienced identification,sources,basic application. Research fronts include cell identifications, liver regeneration, cell differentiation, expression of exogenous genes,etc. Conclusion China should increase the liver stem cell funding, strengthen cooperation with the top institutes, independent innovation, leading liver stem cells research from basic research to clinical research.
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Objective To explore the in vitro isolation of ?2m-/Thy-1+ bone marrow-derived liver stem cells(BDLSCs) which bear double features of stem and liver cells from bone marrow stem cells(BMSCs)as so to provide suitable donor cells for the treatment of liver diseases by cellular transplant. Methods ?2m-/Thy-1+ BDLSCs were isolated by magnetic bead cell sorting(MACS) method from cholestatic rats in vitro,and cell purity was detected using flow cytometry.Liver associated phenotype markers were characterized by RT-PCR and immunofluorescence staining. Results BDLSCs isolated by MACS were purified and viable,and possessed hepatocyte-like features at gene and protein levels. Conclusion ?2m-/Thy-1+ BDLSCs are special subsets of BMSCs which may have promising potentials in the stem cell-based treatment of liver diseases.
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Objective To study the function of liver stem cells during at liver regeneration(LR) at transcriptional level.Methods The liver stem cell growth-and differentiation-related genes were obtained by data collection and literature review,and the gene expressions in rat regenerating liver were checked by Rat Genome 230 2.0 array.The relativity of the LR-associated genes was identified by comparing the discrepancy of gene expression changes between partial hepatectomy(PH) group and sham operation(SO) group.Results Fifty genes were found to be related with liver regeneration.The initial and total expressing gene numbers occurring in the initiation phase of liver regeneration(0.5-4 hours after PH),G_0/G_1 transition(4-6 hours after PH),cell proliferation(6-66 hours after PH),cell differentiation and structure-function reconstruction(72-168 hours after PH) were 24,10,21,2 and 24,23,46,26 respectively,indicating that the related genes were mainly triggered during the initiation phase,and worked at different phases.The total frequencies of their up-and down-regulation expression were 153 and 123 respectively,demonstrating that the expression of the major genes increased,and the minority decreased.Their expression profiles were classified into 6 types,indicating that the activities mentioned above were diverse and complicated during liver regeneration.Conclusion The growth and differentiation of liver stem cells are enhanced during liver regeneration.