动物造模,原代小肠上皮细胞模型 z-bo 2021-10-14 371

　　Objective: To study the proliferation characteristics of primary tree intestinal epithelial cells, and to establish a cell model of human rotatable G1P [8] virus infection in tree primary intestinal epithelial cells.

　　Method: By digesting the combination of collagenase XI and neutral protease I, primary small intestinal epithelial cells of the small intestine of the tree were obtained. After purification and identification, the cells were infected with human spinner G1P [8] virus, and the virus titer of the culture supernatant was measured. The expression of human rotavirus G1P [8] type VP6 protein was detected by Westernblot and indirect immunofluorescence detection, and the infectivity of human rotavirus G1P [8] type virus to primary small intestinal epithelial cells was evaluated in vitro. finished.

　　Result: The isolated primary small intestinal epithelial cells of the tree small intestine were purified by subculture, and the purity of the obtained primary small intestinal epithelial cells of the branch was 90%. The rotavirus susceptibility of primary small intestinal epithelial cells, primary kidney cells, HCT116 and MA104 cells were compared, and it was determined that human rotavirus G1P can infect primary small intestinal epithelial cells of tree branches [8]. ]. After 72 hours of culture, the virus titer can reach 2.0 x 105 TCID 50/mL. Western blot and indirect immunofluorescence showed that the expression and distribution of rotavirus VP6 protein can be detected in the small intestinal epithelial cells of grapevines infected with rotavirus G1P [8] virus for 1-5 days.

　　 Conclusion: A method to isolate, purify and culture primary small intestinal epithelial cells of tree sh was established, and an in vitro model of human rotavirus G1P [8] virus infection of tree sh primary small intestinal epithelial cells was established.