中国农业大学学报2017，Vol.22Issue(10)：91-96,6.DOI:10.11841/j.issn.1007-4333.2017.10.10
山羊小肠上皮细胞分离培养与鉴定
Isolation culture and identification of goat intestinal epithelial cells
摘要
Abstract
The aim of this study was to establish a primary goat intestinal epithelial cell culture model in vitro and investigate goat intestinal nutrient absorption,immune regulation and relationship between enteric microorganism and intestinal epithelial cells.Intestinal tissue pieces were used to obtain goat intestinal epithelial cells in vitro.The cells isolated were further separated and purified by single clone method.These cells isolated were then identified by cell morphology,cytokeratin 18 immunofluorescence,vimentin,desmin and cell proliferation curve.The results showed that goat intestinal epithelial cells was successfully cultured and passaged generations 10 by tissue adherent method;No expressions of vimentin and desmin genes in goat intestinal epithelial cells were detected using RT-PCR;Positive reaction of goat intestinal epithelial cells against cytokeratine 18 antigen,green fluorescent under fluorescence microscope was discovered.The eighth generation cells still displayed characteristics of epithelial cells.To the tenth goat intestinal epithelial cells cytoplasm became bigger and started to senesce.In conclusions,the cells isolated were successfully passaged generation 10 by using the tissue adherent method,which provided cell model in vitro for understanding the mechanisms of nutrient substance absorption and regulation in goat intestinal epithelial cells.关键词
山羊/小肠上皮细胞/细胞角蛋白18/波形蛋白/肌间线蛋白/细胞模型Key words
goat/intestinal epithelial cell/cytokeratin 18/vimentin/desmin/cell model分类
农业科技引用本文复制引用
李小芬,詹康,张响英,陶勇,杨晓志,顾文婕..山羊小肠上皮细胞分离培养与鉴定[J].中国农业大学学报,2017,22(10):91-96,6.基金项目
江苏高校品牌专业建设工程资助项目(PPZY2015A083) （PPZY2015A083）
