Safety Evaluation And Quality Control Of Mscs From Hepatitis B Patient In Vitro Expansion The Feasibility To Crue Liver Disease By Transplanting Stem Cell

Objective:To get MSCs from hepatitis B patient,and to valuate their safety after long term culture in vitro,control the quality for the use of transplantation.Methods:The cells obtained directly from bone marrow and culrured in Mesen Pro medium supplemented with FGF,and the morphology of MSCs was observed. Surface antigens of the MSCs were analyzed by the flow-cytometer,Detection the becteria,virus,endotoxin and residual serum of cell suspension,To study the effect of human MSCs on perpheral blood T lymphocytes,The MSCs and lymphocytes were co-cultured in 48 well plates for 72h,T lymphocyte proliferation was measured by using MTT reduction method and the effect of MSCs on the T lymphocyte transformation stimulated by PHA was observed.The oncogenicity of MSCs was verified by the tumorigenesis test in sofo agar.The genetic stability of MSCs was examined by karyotype analysis. To investigate the effects and mechanism of bone marrow mesenchymal stem cells (BMSC) on liver tumors by culturing in BMSC conditioned mediumResult:The MSCs from hepatitis B patient could be passaged to many generations and had strong abilities of proliferation,They expressed stem cell-surface antigens and maintained normal karyotype,to prevent the pollution of bacteria and viruses,They can inhibit the immune response of allogenic T lymphocytes,the rate of inhibition is 45%(P=0.00).but there was not oncogenicity. Hepatoma cells were seeded in BMSC conditioned medium.The effect were observed under fluorescence microscope to identify BMSC within the histopathological sectionConclusion:The MSCs have proliferative potential,they can be passaged in long-term cultures in Mesen Pro medium without oncogenicity,and can maintain normal karyotype,and they also can inhibit the immune response of T lymphocytes and alleviate the grafe-versus diseases.The MSCs can be served as a news type of cells in cell and gene therapy.But can accelarate cell proliferation by direct contact or through soluble cytokines.