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[Objective] To detect the L-02 liver cell function in embryonic stem cell lines for acute liver failure in rats in order to verify its role in the treatment of acute liver failure, looking for hepatocyte transplantation and bioartificial liver (BIOARTIFICIAL LIVER, BAL) A new hepatocytes available materials. [Methods] 1. Taken the logarithmic growth phase and the proliferation of good L-02 cell lines were extracted RNA and protein, using RT-PCR and of WESTERN-BLOTTING from the L-02 cell lines albumin and a variety of gene and protein level detection liver cell-specific expression of the enzyme. Immunofluorescence labeled albumin and glucuronic acid transferase enzyme, laser scanning confocal microscope under direct observation of the expression. 2 take L-02 cells grow well, adjust cell count to 5 * 10 7/ML the 1ML cell suspension by spleen implanted in rats in vivo (N = 10, G1), and transplanted in 24 hours after the line rats with 90% hepatectomy-induced acute liver failure, observe the protective effect of L-02 cells from rats with acute liver failure. Infusion of cells acute liver failure in rats as a control (N = 15, G2). Blood every 24 hours once the model is established, changes in blood ammonia and liver function parameters were measured, and statistical survival. 3 Take 2 * 10 6 L-02 cells were implanted into nude shoulder, and at the same time take the same amount HEPG-2 cells implanted in the same nude mice contralateral shoulder, comparison of tumor was observed. 【Results】 1 RT-PCR detected L-02 cells expressing albumin (ALB), glutathione-S-transferase (GST-Π), coagulation factor X (HBCF-X). WESTERN-BLOTTING detect visible albumin bands, and the amount of albumin expression similar to human primary hepatocytes. Immunofluorescence confocal laser scanning microscope observed expression of albumin and glucuronic acid group transferase (UGT). 2 with the control group of acute liver failure in rats compared infusion of L-02 cells of acute liver failure in rats significantly longer survival time after infusion of cells in rats week survival rate of more than 70%, while the control group rats all died within 72 hours. The experimental rats hyperammonemia and hepatic function was significantly improved, within 24 hours after hepatectomy elevated, declining more than 48 hours. Control rats continued to rise, and the 24-hour rise higher than the experimental group. Two groups of nude mice tumor: HEPG-2 cells were injected two weeks after the observed tumor growth, L-02 cells without tumor. [Conclusion] The above experiments show that L-02 cells with some synthetic and secretory function of the normal human liver cells to express albumin and a variety of specific enzymes of the liver cells, and that of L-02 cells is not out of the nature of the cells of the liver. L-02 has a longer survival time in rats with acute liver failure, improve liver function, and without the risk of tumor formation through animal experiments further confirmed. In short, with the human liver cell function and without a dangerous tumor of the L-02 cell lines, is expected to provide a better source of cells for liver cell transplantation and BAL.
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