| Liver transplantation (LT) is the only currently effective therapy for end-stage liver disease and severe acute liver failure, but its use is limited by high cost and a shortage of allografts. Therefore, researchers are focusing on differentiation of human stem cells towards hepatocytes as an alternative cell source for liver-based cell therapies.Human adipose-derived stem cells (hADSCs) have been proved to be an attractive candidate for autologous cell transplantation source for regenerative medicine. They can be obtained repeatedly, in large quantities, and in a more convenient, safe, and less painful way. Recently, ADSCs were reported to be able to differentiate into hepatocytes both in vitro and in vivo. Therefore, ADSCs represent a potential source of autologous adult stem cells for cell therapy of liver diseases.Caveolae are distinct domains of the plasma membrane of most cells, where cellular processes such as signaling and membrane sorting occur in a highly regulated lipid and protein environment. Caveolin-1is an essential component of caveolae and has been previously implicated in the regulation of cellular proliferation and lipogenesis.Aim:To investigate the mechanisms of the hepatogenic induced stem cells, we have established the hepatocyte-like cell mode by co-inducing the adipose mesenchymal stem cells with growth factors and discussed the role and signal pathway of Cav-1during the diffrentiation of hepatocyte-like cell by detecting the expression of Cav-1during the process of stem cell inducing into hepatocyt-like cells.Methods:1, hADSCs phenotype was identified by flow cytometry after separating, extracting and culturing the human adipose mesenchymal stem cells.2, hADSCs were induced with HGF, FGF, EGF and dexamethasone referring to the induction methods of R. Talens-Visconti. The induced cells were identified by Immunofluorescence and RT-PCR.3, The safety and effectiveness of magnetic iron oxide nanoparticle in vivo MR imaging tracking were evaluated by in vitro NMR.4, Western blot studies were carried out to detect the MAPK signaling pathway and expression level of Cav-1, ER-a36.Results: 1. After21days of osteogenic induction, the shape of hADSCs changed, mineralized nodules were stained by Von kussa Induced3-4days, hADSCs assumed a rounded shape. Lipid-filled cells were apparent. Intracellular droplets were stained red by Oil Red O, confirming the presence of lipid and indicating their potential to differentiate along an adipogenic lineage. We cultured hADSCs has strong Proliferation ability, differentiation ability.2. Established the hepatocyte-like model with growth factors induced. With the time Oct4, Nanog expression decreased gradually; ALB, CYP1A1, HNF1A, HNF1B gene expression amount increases gradually; AFP expression increased firstly and then decreased.3. Superparamagnetic iron oxide nanoparticles labeled adipose-derived stem cells. At25μg/mL concentration cell proliferation levels were normal, and in MR detection were significant differences.4. Cav-1and ER-a36expression with induction time was significantly increased. After7days MAPK signaling pathway is activated.Conclusion:The sufficient number of adipose mesenchymal stem cells which was extracted from adipose tissue can amplify steadily and sustainably in vitro. It has multiple differentiation ability. The detecting of associated protein and genes confirmed the successful of this model. The MAPK signaling pathway mediated the differentiation process of serum cells to hepatocyte-like cells in which Cav-1played an important role. Our findings may provide the foundation for the investigation of inducing and differentiation mechanisms of stem sells to hepatocyte-like cells. |
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