动物造模,原代肾上皮细胞感染模型 zi-bio 2024-03-06 640

　　Objective: To establish an EV71 infection model on primary kidney cells of tree shrews.

　　Method: Trypsin digestion was used to obtain primary kidney cells from tree shrews. EV71 was used to infect tree shrew kidney cells, and the viral titers of the supernatant cultured at 1, 2, 4, 6, and 8 days were measured. Western blot and indirect immunofluorescence were used to detect the expression of EV71 virus VP1 protein in the cells, respectively, to determine the infectivity of EV71 virus to tree shrew primary kidney cells.

　　Result: The isolated primary kidney cells of tree shrews were passaged, purified, and morphologically differentiated, and a cell culture mainly composed of tree shrew primary kidney cells was established. Infection of tree shrew primary kidney cells with EV71 virus resulted in a viral titer of 1.3 × 106 TCID50/mL at 48-96 hours after infection, indicating that EV71 virus can effectively infect and proliferate tree shrew primary kidney cells. Western blot analysis revealed that the VP1 protein of EV71 virus can be effectively detected in primary kidney cells of tree shrews 2-8 days after infection. Indirect immunofluorescence assay detected the distribution of VP1 protein in the cytoplasm of cells 2-6 days after infection.

　　Conclusion: Based on the successful establishment of primary kidney cell culture in tree shrews, the infectivity and viral proliferation characteristics of EV71 virus on tree shrew primary kidney cells were determined, and a preliminary EV71 tree shrew primary kidney cell infection model was established.