动物造模,骨髓间充质干细胞 z-bio 2022-02-16 520

　　Objective: To observe the effects of lentiviral vector-mediated HCV receptor gene OCLN/CD81 transfection on the biological characteristics and gene expression of tree shrew bone marrow mesenchymal stem cells under in vitro culture conditions.

　　METHODS: Tree shrew BM-MSCs were isolated and identified in our laboratory, lentiviral vectors containing human OCLN/CD81 gene were constructed, and CD81 and OCLN genes were transfected into BM-MSCs, respectively. The transfection efficiency was detected, the cell proliferation of BM-MSCs after transfection was detected by CCK8 method, and the multi-directional differentiation potential of osteoblasts was detected by induced differentiation of osteoblasts. The HCV receptor (OC/CD81) gene was detected by real-time fluorescence quantitative and WB methods. mRNA and protein expression and expression of stemness genes in BM-MSCs after transfection.

　　RESULTS: After transfecting the lentiviral vector containing human OCLN/CD81 gene at MOI of 100, the transfection efficiency of LV-CD81 was 99.4%, and the transfection efficiency of LVOCLN was 22.6%. The cell proliferation trend was similar to that of BM-MSCs without gene transfection. , the transfected BM-MSCs could still differentiate into osteoblastic adipocytes, but the ability was decreased. The expression of stemness gene NANOG mRNA was increased, and the difference was significant (P<0.05), and the expression of LIN28A mRNA was decreased, and the difference was significant. (P<0.05). After transfection, tree shrew BM-MSCs can express the transfected exogenous genes efficiently.

　　Conclusion: The constructed lentiviral vector containing human OCLN/CD81 gene can successfully transfect tree shrew BM-MSCs and express the transfected gene efficiently. Influence.