Rat Bone Marrow Mesenchymal Stem Cell Inducation And Intrahepatic Transplantation

Chronic hepatitisf portal hypertensionf liver fibrosisf primary hepaticcancerf hereditary and metabolic hepaticdisease are difficult problem front allover the world. Now the most effective treaty method is Original livertransplantation. But lack of donorf immunological rejection f very expensiveand others things limit its apply. In recent years hepatocyte transplantatHTx have become a new treatment of various liver failure. Hepatocyte cankeeping surving for a long time in hetro-location and play the function ofhepatocyte on a certain degree.A little amount of transplanted hepatocytemay obtain some purposes by proliferating in vivo such as rectifying thecongenital hepatic metabolic obstacle supporting liver function temporarilyand substituting injured liver tissue etc. HTx technique show a promisingapplied foreground gradually along with the further development of cellbiology and growth biology technique. However there are still many problemsrequired to be resolve today such as lack of hepatocytes source f insufficientof the number of transplanted cells f cells proliferation after transplantation fsubstitution degree of injured liver tissue f supporting time of function andimmune rejection etc. Although great effort have been made all over theworld the effect is not very good so the clinic application of HTx isinfluenced. Therefore it is urgent to look for a new treatment method.Stem cells are natural units of the proliferation of embryo cells and theregeneration of mature cells and all kinds of tissue .Stem cells can not onlyproliferate powerfully but also differentiate to all kinds of celltype .Transplanted stem cells can not only replace injured tissue but alsostimulated receptor tissue proliferating and repairing itself. In recent yearsresearch on mesemchymal stem cells MSCs plasticity have made greatprogress.MSCs are pluripotent cells and can differentiate intonon-hematopoietic cell type under certain conditions such as osteoblastfchondroblastsf adipocyte skeletal myoblastsf neurocytesf and neuroglial cells.After being transplanted MSCs can differentiate to albumin-excretinghepatocyte in vivo so that MSCs provide new donor source for curing hepaticdisease by HTx. MSC shave higher proliferation ability angd lower immunitycharacter compared with mature hepatocytes Clinic application foreground ofMSCs is very outstanding . because it not only have advantage of being cellssource of hepatocytes transplantation but also avoid the ethnics debate onembryo stem cell research.[OBJECTIVES]The aim of this study is to reseach on bone marrow MSCs isolating fpurified method observe the inducing effect of injured liver tissuehomogenate and bone marrow MSCs growth character from cellmorphology then transplanting the cells into the liver of emergency liverfailure rat. We detect the transplanted cells function by serum biochemicalindex so that we obtain the evidence that MSCs cure hepatic disease byintrahepatic transplantati0on .[METHOD]Isolated and purified MSCs fronm bone marrow of young rat by densitygradiend centrifugation and adhering to the culture plastic.After successivesubculture and amplification the growth curve were drawn the morphologywere observed under microscope.Passage 3 were induced in 10%FBS f 10%injured liver homogenate in vitro observed MSCs morphological changeand differentiated character detectived albumin byimmunocytochemistry .Then transplanted the cell into emergency liver failurerat by intrahepatic injection and detective rat serum biochemical index atdifferent time g[RESULT]1f The isolated MSCs were mononuclear cells in bone marrowtheirliving behaviores was quite stable in L-DMEM containing 10% FBS andthe growth curves of passage 1f 3f 5were auite similar exhibiting a largeexpensive potential and the typical fibroblast-like morphology.2f Passage 3 differentiated to adipocyte in L-DMEM containing 20%horse serum Red O staining result was positive.