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用携带CTLA4Ig基因的重组腺病毒感染大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs),体外向肝细胞诱导分化,并检测其免疫抑制功能.用含有HGF等细胞因子培养液诱导重组腺病毒Ad-CTLA4Ig感染大鼠BMMSCs向肝细胞分化.诱导后的细胞可表达AFP、Alb和CK18等肝细胞标志,同时还具有储存糖原和摄取、排泌靛青绿等肝细胞功能.转基因BMMSCs在未经诱导和诱导后均可表达CTLA4Ig,诱导14天时表达量有所减弱.单向混合淋巴细胞反应证实,诱导7天的转基因BMMSCs具有明显的抑制淋巴细胞反应的作用,其抑制作用明显高于未转基因BMMSCs,且CTLA4Ig基因修饰BMMSCs输注还可以明显延长肝移植大鼠的存活时间.用重组腺病毒Ad-CTLA4Ig对BMMSCs进行基因修饰,一方面不会影响BMMSCs的肝细胞分化潜能,另一方面使BMMSCs的免疫抑制特性得到进一步强化.
Rat bone marrow mesenchymal stem cells (BMMSCs) were infected with the recombinant adenovirus carrying CTLA4Ig gene and induced to differentiate into hepatocytes in vitro, and the immunosuppressive function of the cells was assayed.The recombinant cells were induced by recombinant cytokines such as HGF Ad-CTLA4Ig adenovirus Ad-CTLA4Ig infected BMMSCs differentiate into hepatocytes.The induced cells can express hepatocyte markers such as AFP, Alb and CK18, and also have the function of storing glycogen, uptake and excretion of indocyanine green, etc. Transgenic BMMSCs The expression of CTLA4Ig could be expressed in both untransformed and induced groups, and the expression of CTLA4Ig was attenuated on the 14th day.It was confirmed by one-way mixed lymphocyte reaction that the transgenic BMMSCs induced for 7 days had obvious inhibitory effect on lymphocyte reaction and the inhibitory effect was obvious In non-transgenic BMMSCs, and CTLA4Ig gene modified BMMSCs infusion can also significantly prolong the survival time of liver transplantation rats.Using recombinant adenovirus Ad-CTLA4Ig gene modification of BMMSCs, on the one hand will not affect the BMMSCs hepatocyte differentiation potential, and the other On one hand, the immunosuppressive properties of BMMSCs are further strengthened.