Preliminary Studies On Transplantation Mesenchymal Stem Cells To Treat Rat Fulminant Liver Failure And The Mechanism Of The Effect

PartⅠSeparation,cultivation and identification of rat mesenchymal stem cellPurpose Separation , cultivation and purification of rats mesenchymal stem cells; Using the flow cytometry technology to judge whether the cell is MSCs.Method Apply the screening method of adherent culture of whole bone marrow cell to separate and purify the rat MSCs. Analysis the cells differentiation phenotype during the process of cultivation to judge whether the cells is MSCs.Result The rat MSCs looks fusiform under light microscope.Useing the flow cytometry to detect the cell phenotype,the result is that the expression of CD90.1 and CD29 is high(99.2% and 98.9%),the expression of CD34 and CD45 is low(0.5% and 0.9%). It is speculated that the cultured cells is MSCs.Conclusion The rat MSCs can be succesfully isolated and cultured by whole bone marrow adherent culture. PartⅡTransplantation mesenchymal stem cell to treat rat fulminant liver failurePurpose Transplant the cells into the rat model of acute liver failure induced by CCL4,then compare the liver function,survival rate, the homing efficiency , the pathological changes and the level of TNF-αamong the model control group and the normal control group.Method MSCs cultivated in vitro as the first part and were labeled by DAPI. Establish the rat acute liver failure model induced by CCL4, Divided the rats into two groups: Experimental group and model control group,the rats of normal control group intragastric administrate the same quantity saline. labeled bone marrow stem cells were suspended in 1.5ml saline at a concentration of 1×106 cells and transplanted by tail vein injection to experimental group and normal control group,the model control group administrate the same quantity saline by tail vein injection.Comprare the experimental group and model control group,Observe the general situation and survival rate of two group rats, survival rats were executed on 7d and 14d after transplantation, Blood samples were taken from the rats, detect the liver function, TNF alpha level,and the liver tissues were sectioned into slices,observe the liver pathology change, used Frozen section analysis under fluorescence microscope to detect the MSCs in the liver parenchyma, and compared with the normal control group.Result About 98% of the cells were successfully labeled by DAPI. The rats Liver failure was established induced by CCL4, The liver failure was characterized with decreaed activity,downcast,hypnody,exanimation,low response to stimulus and urinary incontinence,liver function was severely damaged,and the histological examination shows hepatocyte necrosis along with the inflammatory cell infiltrate.All the above have aproved that the establishment of the rat model of acute liver failure induced by CCL4 is successful. There were significantly increase in the general situation on 3d in Experimental group when compared to model control group. On 7d and 14d after transplation ,Experimental group has a better improvement of liver function, survival rate, TNF-αlevel, liver pathology change than model control group.The Experimental group could find DAPI labeled cells on 7d and 14d after transplation.The model control group could not find DAPI labeled cells on 7d and 14d after transplation.The normal control group could find several DAPI labeled cells on 7d and 14d after transplation.Conclusion Bone marrow mesenchymal stem cells (MSC) can home to liver failure rat liver, can promote the acute liver failure rat liver function, reduce the liver tissue necrosis and inflammation, promote liver immunifaction to repair the tissue, has obvious curative effect for acute liver failure rat.