动物造模,人轮状病毒G1P[8] z-bio 2021-12-15 282

　　OBJECTIVE: To investigate the proliferation characteristics of primary small intestinal epithelial cells of tree shrews and establish an in vitro cell model of human rotatable G1P [8] virus infection of primary small intestinal epithelia of tree shrews.

　　Method: Through the combined digestion of collagenase XI and neutral protease I, primary small intestinal epithelial cells of the tree sh are obtained. After purification and identification, the cells were infected with human rotatable G1P [8] virus and virus titer. The expression of human rotavirus G1P [8] type VP6 protein was detected by Western blotting and indirect immunofluorescence detection, and the infectivity of human rotavirus G1P [8] type virus to Tupai primary small intestinal epithelial cells was determined. It was evaluated in vitro.

　　Result: The isolated primary small intestinal epithelial cells of the tree sh were purified by subculture, and the primary small intestine epithelial cells of the tree sh with a purity of 90% were obtained. The rotavirus susceptibility of Tupapa’s primary small intestinal epithelial cells, primary kidney cells, HCT116 cells and MA104 cells were compared, and it was determined that Tupapa’s primary small intestinal epithelial cells could be protected by human rotavirus G1P[8] Infection. After 72 years, the virus titer can reach 2.0×105 TCID 50/mL in the culture time. Western blot and indirect immunofluorescence showed that the expression and distribution of rotavirus VP6 protein can be detected in Tupa human small intestinal epithelial cells infected with rotavirus G1P [8] virus for 1-5 days.

　　Conclusion: A method for isolating, purifying and culturing tree sh primary small intestinal epithelial cells has been established, and an in vitro model of human rotatable G1P [8] virus infection of tree primary small intestinal epithelial cells has been established.