Study On The Characteristics Of Human Amnion-derived Stem Cells And Their Potential Therapeutic Effect On Experimental Hepatic Fibrosis

The use of stem cells to generate biological substitutes and improve functions for damaged tissue/organ with high proliferability and differentiability has been the most attractive focus in regenerative medicine nowadays. Recently, amnion-derived cells , which contains amniotic epithelial cells and amniotic mesenchymal stem cells,have been reported to have multipotent differentiation ability, and these cells have attracted attention as a cell source for cell-transplantation therapy. The amnion possesses considerable advantageous characteristics: the isolated cells can differentiate into all three germ layers; they have low immunogenicity and anti-inflammatory functions; and they do not require the sacrifice of human embryos for their isolation, thus avoiding the current controversies associated with the use of human embryonic stem cells. Therefore, amniotic membrane has been proposed as a good candidate to be used in cellular therapy and regenerative medicineIn this study, we intended to investigate the multilineage differentiation abilities of these two types of amnion-derived stem cells and their potential therapeutic effect on hepatic fibrosis.AIM:To investigate the multipotent differentiation ability of human amniotic epithelial cells (hAECs) and human amniotic mesenchymal stromal cells(hAMSCs); And to evaluate the feasibility of transplanting these two types of cells in treatment of liver fibrosis on mouse model. .METHODS:hAECs were obtained by subsequent trypsin digestion after elective cesarean section of healthy pregnancy women at term, while hAMSCs must go to next digestion by collagenase. These two types of cells were induced to differentiate into osteogenic, adipogenic, chondrogenic, and hepatic cells in vitro. The differentiation potential of hAECs and hAMSC were evaluated by flow cytometry, specific staining, mRNA expression and immunocytochemical assay.We also investigated the therapeutic effect of hAECs and hAMSCs on carbon tetrachloride (CCl4)–induced liver fibrosis in mice. Immune-competent C57/BL6 mice were treated with CCl4 by intraperitoneal injection for 4 weeks, and then followed by a direct injection of hAECs and hAMSCs, respectively, through tail vein. PBS only as a control. After 2 more weeks, transplanting groups with hAEC and hAMSC , and control group with PBS were examined by Masson staining ,ELISA assay of serum albumin levels and ultrasonic scanning. RESULTS:hAECs and hAMSCs had differentiated into cells of osteogenic, adipogenic and chondrogenic in culture supplemented with certain inducing factors in vitro. Transplantation of hAECs or hAMSCs into mouse model of CCl4–induced liver fibrosis leads to survivals of xenograft-cells, reduced hepatic fibrosis in comparison with mice without hAEC or hAMSC transplant.CONCLUTION:Both hAECs and hAMSCs have the potential to differentiate into various types of cells, including osteogenic, adipogenic, chondrogenic, and hepatic cells in vitro. hAEC or hAMSC transplantation effectively reduces experimental hepatic fibrosis, providing a novel approach for the treatment of fibrotic liver diseases in the near future.