The Study Of The Induction Of Human Induced Pluripotent Stem Cells And Its Differentiate To Hepatocyte Like Cells In Vitro

Background:At present, the most effective treatment liver failure is liver transplantation. But due to the severe shortage of donor,liver transplantation can not be popularized. Artificial liver support system can replace the liver function temporarily, help patients through liver failure risk for the regeneration of liver cells, or to maintain the survival to get suitable donors for liver transplantation. Biological artificial liver is currently considered the most ideal artificial liver, but its development is limited by the origin problem of liver cells. Stem cells have the ability to organize into various types of cells, may be the new source of liver cells for artificial liver treatment. Embryonic stem cells have good pluripotency, can differentiate into a variety of human tissues and cells, but its application is restricted by ethics. Autologous bone marrow mesenchymal stem cells and peripheral blood stem cells are difficult to obtain enough quantity, not easy to establish a cell line, and difficult to meet the required quantity and degree of stability of biological artificial liver.Induced pluripotent stem cells,like embryonic stem cells, have the totipotent of self-renewal, proliferation and differentiation, but avoid the ethics and law obstacles the embryonic stem cell research had been facing, avoid The main disadvantage of the common dissimilar and tumor liver cells used in bioartificial liver at present, may become a new source of liver cells of the treatment of end-stage liver failure, has broad prospects of clinical application.Aim:In orde to provide a new source of cells for artificial liver support treatment. human foreskin fibroblasts were induced into iPS cells, and further induced into hepatocyte like cells.Method:In this paper, Oct4, Sox2, Klf4 and c-Myc gene were transduced into human foreskin fibroblast by retroviral vector in vitro. The virus infected cells were cultured by hES cell culture system, and were induced into iPS cells.The pluripotency of iPS cells like ES cells were identified by AP staining, immunofluorescence staining, karyotype analysis, embryoid body formation, and umbilical embryo tumour formation. The obtained iPS cells were adding different stimulating factor in vitro. IPS cells were firstly induced differentiation into endoderm cells, and then differentiate to hepatocyte like cells. The process of induced were proved to be correct by immunofluorescence staining and flow cytometry used markers of each stage, the differentiation efficiency is high.Elisa method was used for detection of ALB secretion of the hepatocyte like cells, PAS staining was used to detect glycogen storage function, RT-PCR and Real-time PCR was used to detect the expression of specific gene. IPS cells by fluorescence labeled injected in C57 mice with acute liver failure via tail vein in mice to understand whether the iPS cells have a role in the treatment of liver injury.Result:Human foreskin fibroblasts were success induced to iPS cells, and were identified to have the pluripotency similar to ES cells. The obtained iPS cells in the experiment in vitro were adding different stimulating factor, then the iPS cells were induced into the hepatocyte like cells with hepatocyte function. IPS cells by fluorescence labeled injected in C57 mice with acute liver failure via tail vein in mice had certain role in the recovery of liver function damage.Conclutiom:Human foreskin fibroblasts can be induced into iPS cells, and further induced into hepatocyte like cells, may be the potential source of liver cells for biological artificial liver in treatment of liver failure.