Isolation, Identification Of Fetal Liver Stem Cells And Transplantation For The Treatment Of Acute Liver Injury In Nude Mice

Objective To study the initial biological properties of fetal hepatic progenitor/stem cells,and the feasibility and effectiveness of their transplantation on acute liver injury nude mice.Methods The fetal hepatic progenitor/stem cells isolated from 13.5-14.5 dpc pregnant mouse fetal liver by way of enzyme digesting were cultured in vitro and stem cell specific markers as AFP,CK19,Albumin,c-Met,E-cadherin were identified by immunocytochemistry,immunofluorescence,Westernblot and RT-PCR on colonies,used phase-contrast microsope and transmission electron microscope to observe the cell apperance and inside structure;used glycogen staining to observe expression of glycogen,Flow cytometry was used to determine the cell cycle disposition.At last,4×10~5 cells were transplanted into nude mice with carbon tetrachloride induced acute liver injury.Hapatic functions were measured pre and post transplantation on days 1,2,4,7.Meanwhile,hepatic pathology was studied.Results We cultured stem cell-like liver progenitor cells from mouse fetal liver with positive staining of Albumin,cytokeratin19(CK19),c-Met,AFP,E-cadherin.In experiment of cell transplantation,compared to control group the hepatic functions gradually recovered in transplant group,on days 1,2,4 after fetal hepatic stem cell transplantation.The hepatic pathology significantly improved in stem cells transplantation group.Conclusion Fetal hepatic progenitor/stem cell were successfully yield by enzyme digest,with transplantation stem cells improved the hepactic function and pathology in acute hapatic injury model of nude mice.