动物造模,慢病毒载体介导HCV受体基因 z-bio 2021-12-23 422

　　Objective: To observe the effects of lentiviral vector transfection with HCV receptor gene OCLN/CD81 on the biological characteristics and gene expression of tree bone marrow mesenchymal stem cells in vitro culture?

　　Method: Use the trees separated and preserved in this chamber to harvest from BM-MSC, construct a lentiviral vector containing human OCLN/CD81 gene, and transfect them into BM-MSC respectively, and then detect with a fluorescent inverted microscope and flow cytometer Transfection efficiency. I used the CCK8 method. Have you detected the proliferation of BM-MSC cells after transfection, induced differentiation of osteoblastic adipocytes, and tested multi-directional differentiation potential? Real-time fluorescence quantification and WB method to detect HCV receptor (OC/CD81) gene mRNA and protein expression. What is the gene expression of BM-MSC stem cells after transfection?

　　Result: After transfecting the lentiviral vector containing human OCLN/CD81 gene with MOI100, does the transfection efficiency of LV-CD81 reach 99.4%, and whether the transfection efficiency of LVOCLN reaches 22.6%? The cell proliferation trend is similar to that of untransfected BM-MSC. Transfected BM-MSC can still differentiate into osteoblasts, but is its capacity reduced? The stem gene NANOG mRNA expression increased, the difference was statistically significant (P\u003c0.05), and the LIN28A mRNA expression decreased, the difference was significant (P\u003c0.05)? After transfection, can the stem cell BM-MSC effectively express the translocated foreign gene?

　　Conclusion: The constructed lentiviral vector based on the human OCLN/CD81 gene can successfully transfect Tree Schrew BM-MSC and effectively express the transferred gene. Does transfection have a certain effect on the multi-directional differentiation potential of Tree Schrew BM-MSC?