中国实验动物学报2017，Vol.25Issue(2)：111-116,6.DOI:10.3969/j.issn.1005-4847.2017.02.001
人轮状病毒G1P[8]型感染树鼩原代小肠上皮细胞模型的建立
Establishment of a model of tree shrew primary small intestinal epithelial cells infected with human rotavirus G1P[8]
摘要
Abstract
Objective To explore the proliferation characteristics of primary small intestinal epithelial cells of tree shrews and the characteristics of human rotavirus(RV) G1P[8] infection to these cells,and establish a model of tree shrew primary small intestinal epithelial cells infected with human rotavirus G1P[8].Methods The primary small intestinal epithelial cells were obtained by collagenase Ⅺ and dispase I digestion from tree shrew.After purification and identification,the obtained primary small intestinal epithelial cells were infected with RV.Then,culture supernatants of infected cells were collected every 12 hours after infection.Viral titer and viral load were subsequently determined.Western blot and indirect immunofluorescence observation were used to detect the expression of RV protein VP6 in the primary cells.The infectivity of RV to the tree shrew primary cells was finally evaluated.Results After purification and identification of primary epithelial cells from the tree shrew,high purity above 90% primary tree shrew small intestinal epithelial cells was obtained.These primary small intestinal epithelial cells could be infected with RV virus by comparing the virus infectivity to primary renal cells,HCT116 cells and MA104 cells.The virus titer reached to 2.0×105TCID 50/mL at 72 h after infection.Using Western blot and indirect immunofluorescence observation,the specific viral protein of VP6 was determined to be expressed in the tree shrew primary small intestinal epithelial cells,and were located in the cytoplasm from days 1 to 5.Conclusions The separation,purification and cultivation methods of tree shrew primary small intestinal epithelial cells are successful,and the tree shrew model of RV-infected the tree shrew primary small intestinal epithelial cells is successfully established.关键词
树鼩原代小肠上皮细胞/轮状病毒/病毒增殖特性/VP6蛋白Key words
Primary intestinal epithelial cells/Tree shrews/Rotavirus/Virus proliferation/Protein VP6分类
生物科学引用本文复制引用
李道群,彭杰,甸子芩,王文广,张阿梅,冯悦,牛华,代解杰,夏雪山..人轮状病毒G1P[8]型感染树鼩原代小肠上皮细胞模型的建立[J].中国实验动物学报,2017,25(2):111-116,6.基金项目
国家科技支撑计划项目(编号:2014BAI01B01). （编号:2014BAI01B01）
