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

在体外分离培养奶牛瘤胃上皮细胞，可为奶牛瘤胃生理功能和吸收机制研究和永生化奶牛瘤胃上皮细胞奠定基础。通过胰蛋白酶消化法从奶牛瘤胃组织中分离出瘤胃上皮细胞，并通过免疫细胞化学的方法检测细胞角蛋白，CCK-8检测细胞生长曲线，β-半乳糖苷酶染色鉴定细胞的衰老以及细胞染色体核型分析来鉴定瘤胃上皮细胞。结果表明：1）利用胰蛋白酶消化法可以稳定的培养出瘤胃上皮细胞并且能够在体外传代大约5代；2）通过在显微镜下进行细胞形态的观察呈单层"岛屿状"；3）在荧光显微镜下细胞角蛋白抗原能够发出绿色荧光；4）奶牛瘤胃上皮细胞传至第5代，β-半乳糖苷酶被染成蓝色，同时，细胞出现衰老和停滞生长；5）染色体核型分析表明奶牛瘤胃上皮细胞的染色体数为60条。研究发现通过酶消化奶牛瘤胃组织能够获得奶牛瘤胃上皮细胞，但只能传至5代。
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.