Abstract: Abstract: Objective To isolate intestinal epithelial cells( IECs) from the tree shrew，to establish a cellular model for drug mechanism research，and to lay the foundation for establishment of intestinal epithelial cell lines． Method Collagenase type XI，and neutral protease type I and DTT were used for digestion of tree shrew small intestine． Cells morphology was observed by phase contrast microscopy，identified by cytokeratin Immunofluorescence assay．Ｒesult Isolated intestinal epithelial crypts attachment were performed in 24 h，confluent in 6 d． Epithelial outgrowth from isolated crypts were formed with confluent monolayers cobblestone and multi-angular morphology． Cells were passaged every 2 ～ 3 d from the nest generation，and between the third and sixth day from the day when cells were plated the cells got into the logarithmic growth phase． The cells were stained positive for cytokeratin immunofluorescence staining． Conclusion Intestinal epithelial cells were successfully isolated from the tree shrew， and the method of isolation for tree shrew’s intestinal epithelial cells was established．

Key words: <p>tree shrew, intestinal epithelial cell, enzymatic digestion, Immunofluorescence, identification</p>

摘要： 摘要: 目的 从树鼩小肠分离小肠上皮细胞，探索分离培养方法和条件，为药物开发和病原感染机制研究提供体外细胞模型，并且为建立树鼩小肠上皮细胞系奠定基础。方法 采用胶原酶Ⅺ和中性蛋白酶Ⅰ及 DTT 联合消化树鼩小肠组织块，得到的细胞经完全培养液培养，用相差显微镜观察细胞形态，通过 MTT 法测定细胞生长曲线，并采用细胞角蛋白荧光染色等方法鉴定细胞。结果 经优化分离法得到的肠上皮隐窝细胞团在 24 h 后贴壁，6 d 汇合成片，在显微镜下观察发现细胞团延辐射状向外长出铺路石样和多角状的细胞; 第二代后，每隔 2 ～ 3 d 可传代 1 次; 细胞接种 3 ～ 6 d 为对数生长期; 细胞角蛋白 18 进行免疫荧光鉴定后确定为小肠上皮细胞。结论 本实验成功地从树鼩小肠分离到了小肠上皮细胞，并且建立了树鼩小肠上皮细胞体外培养的方法。

关键词: <, p>, 树鼩, 小肠上皮细胞, 酶消化, 免疫荧光, 鉴定<, /p>