Treatment Of Transplanted Human Amnion Epithelial Cells For Liver Damage In Mice

Background:Recent years,stem cell therapy have become hot research topics,and the choice for kinds of stem cells is a key issue in clinical application.As a subsidiary in childbirth,source of human amniotic tissue is rich.Human amniotic stem cells,especially human amniotic epithelial cells(hAECs),have the potential to differentiate into the three germ layers and express a variety of embryonic stem cell markers.The characteristics of hAECs,no tumorigenicity,rich sources,easily to get,low immunogenicity and no medical ethics limit,are advantages which embryonic stem cells and many adult stem cells do not have.The liver is a vital organ of the human body.Its main functions include metabolism,detoxification,defense,and hematopoiesis.But there is no cure for severe liver disease.Research on hAECs will benefit cell-transplantation for liver diseases and gene therap.Method:Collect hAECs from human amniotic membrane using enzymatic digestion,observe morphology of cells.Draw the growth curves of the second generation and the fifth generation.Detect the expression of keratin 19 in hAECs by immunocytochemistry.Identify genes related to stem cells by RT-PCR.Make the model of liver damage in mice induced by intraperitoneal injection of carbon tetrachloride(CCl4).Transplant the CFSE-labeled hAECs into the liver damage mice by tail vein injection.Detect the liver function and histopathological changes in mice after transplantation.Result:The hAECs have large nucleus,shapes like pavement and could be continuously subcultured in vitro.Immunocytochemistiy assays showed that hAECs expressed epithelial cell marker keratin 19.RT-PCR assays showed that hAECs express stem cell marker genes REX1,Oct-4,nanog.The hAECs could be induced into adipocytes,neurocytes and liver in vitro and expressed markers which associated to the induced cells,such as PPARγ2,MAP-2,albumin.Frozen sections of immunoflrorescence showed that hAECs could be moved to damaged liver.The repair effect was remarkable in liver function and histopathology after transplanted hAECs into liver damaged mice.Conclusion:Human amniotic epithelial cells can be used for xcmotransplantation and it can promote the recovery of liver injury induced by carbon tetrachloride in mice.