ABSTRACT
AIM: The ?-catenin is a key molecule in the Wnt signal pathway, which plays a critical role in normal development and tumorigenesis. However, the mechanisms of the ?-catenin on the cell growth control are still not completely defined. The aim of this study was to test the hypothesis that the mutant ?-catenin may regulate the hepatocyte proliferation. METHODS: The immortalized murine hepatocyte cell line, AML12, was used for this study. A plasmid that contain mutant ?-catenin S33Y was transfected into the AML12 cells and a stable cell line AML12S33Y was established. The cell growth property of this cell line and the parental cell were compared by flow cytometry analysis and direct cell count. The cells were also tested for the ability to form soft agar colonies, and the ability to form tumors in the severe immune deficient mice (SCID). RESULTS: 1. The mutant ?-catenin containing cell line AML12S33Y has higher proliferating index compared with the parental AML12 cells ( P
ABSTRACT
AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro . METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.
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Objective To establish a recombinant eukaryotic expression vector containing human interferon-?(IFN-?) cDNA,and investigate the expression of IFN-? gene in transfected hepatocellular carcinoma (HCC) cell lines. Methods pcDNA3-IFN-?,established by subcloning IFN-? cDNA into a mammalian expression vector pcDNA3,was mediated into HCC cell lines SMMC-7721 and QGY-7701 by lipofectamine. The transfected HCC cells were selected in RPMI1640 containing G418(400～700?g).RT-PCR analysis and ELISA assay were used for mRNA transcription and protein expression of IFN-? gene. Results IFN-? mRNA was detected only in pcDNA3-IFN-? transfected cell lines, and high levels of IFN-? protein were detected only in the supernatants of IFN-? gene-modified cell lines. Conclusions pcDNA3-IFN-? can stably expressIFN-? cDNA in the transfected HCC cell strains
ABSTRACT
To study the effects of 70%Hepatectomy on the proliferation of rat fetal hepatocytes after intrasplenical transplantation, fetal hepatocytes isolated from 3 week SD rat fetuses bred were transplanted into the spleens of liver regeneration model rats with 70% partial hepatectomy. The cell cycle of hepatocytes in the remanent liver was analyzed by flow cytometer and the density dimensions of the donor fetal hepatocytes in spleen were measured by Image Analysis System 7 and 30 days post transplantation respectively. Compared with the control group, the proportions of S state cells in the remanent liver were obviously increased ( P