中国农业大学学报2018，Vol.23Issue(1)：80-86,7.DOI:10.11841/j.issn.1007-4333.2018.01.10
奶牛瘤胃上皮细胞分离培养与鉴定
Culture and identification of the bovine rumen epithelial cell line
摘要
Abstract
The aim of this study was to isolate bovine rumen epithelial cells in vitro to lay foundation for,the absorption of rumen physiological function for bovine mechanism research and rumen epithelial cells immortalized cows.Bovine rumen epithelial cells were isolated from the rumen tissue by trypsin digestion,and identification of cell protein were identified by immunocytochemistry test,cell growth curve of CCK-8 cell angle was detected,by β-galactosidase staining and to identify the rumen epithelial cells were identified by aging cell karyotype analysis.The results showed:1) Rumen epithelial cells could be cultured stably by trypsin digestion method and used for about 5 generations in vitro;2) Observation of the morphology of cells under the microscope revealed that the monolayer was "Island shaped";3) Fluorescent protein emitted green fluorescence under fluorescence microscope;4) The rumen epithelial cells transferred to fifth generation were β-galactosidase dye blue,at the same time,cell senescence and stagnant growth occurred;5) Karyotype analysis showed that the number of chromosomes in the rumen epithelial cells was 60.In summary,rumen epithelial cells tissue could be obtained through enzymatic digestion,however the cells could only be transferred to fifth generation.关键词
奶牛瘤胃上皮细胞/永生化/细胞角蛋白18/染色体核型Key words
bovine rumen epithelial cells/immortalize/cytokeratin 18/karyotype分类
生物科学引用本文复制引用
姜茂成,詹康,严康,赵倩明,赵国琦..奶牛瘤胃上皮细胞分离培养与鉴定[J].中国农业大学学报,2018,23(1):80-86,7.基金项目
现代农业奶牛产业技术体系专项资金资助(CARS-36) （CARS-36）
国家自然科学基金项目(31572430) （31572430）
江苏省研究生科研与实践创新计划项目(SJCX17_0630) （SJCX17_0630）
农业三新工程(SXGC[2016]326) （SXGC[2016]326）
