中国驯鹿瘤胃上皮细胞的体外分离培养  

The in vitro isolation and culture of rumen epithelial cells in Chinese reindeer

作　　者：范燕茹[1] 王佩[1] 金鑫[1] 王冠玉 邱凯 王秀芳 苏日古格 余兴邦 杨银凤[1] 

机构地区：[1]内蒙古农业大学兽医学院,呼和浩特010018 [2]内蒙古呼伦贝尔市动物疫病预防控制中心,内蒙古呼伦贝尔021008

出　　处：《黑龙江畜牧兽医》2014年第6期8-10,15,228,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31160491)

摘　　要：为了建立驯鹿瘤胃上皮细胞的体外分离及培养方法，试验将驯鹿瘤胃组织进行钝性分离得到上皮层，用0．25％Trypsin-0．02％EDTA消化液37℃连续振荡消化上皮组织3～4h，每30min更换新的消化液，从消化第4次起过滤消化液，离心收集细胞，采用含有15％FBS的M199培养液培养和传代，在显微镜下观察其细胞形态进行形态学鉴定以及采取免疫细胞化学的方法检测细胞内的角蛋白CKl9进行化学鉴定。结果表明：用上述试验方法可以获得能够用于培养的细胞，并能将这些细胞成功地进行了原代和传一代的体外培养。通过细胞形态学观察和免疫细胞化学检测，证实所培养的细胞为上皮细胞。说明试验基本建立了驯鹿瘤胃上皮细胞在体外分离培养的方法。To establish the in vitro melhods for the isolation and culture of rumen epithelial cells in Chinese reindeer. The rumen tissue of the reindeer was bluntly isolated to obtain the epithelial layer, and then the epithelial tissues were treated with 0.25% Trypsin - 0.02% EDTA di- gestive juice under continuous oscillation for 3 - 4 h at 37 ℃. The digestive juice was replaced every 30 minutes. The digestive juice was filtered from the 4th time. The ceils were collected by centrifugation and then were resuspended in M199 medium with 15% FBS for culture and subcul- ture. The morphology of the cells was observed under the microscope for morphological identification, and then the immunocytochemistry method was used to determine cytokeratin -19 （CK19） in the cells for chemical identification. The results showed that the cultured cells could be ob- tained by the above - mentioned test methods, and the ceils could be successfully cultured in vitro for primary culture and the first generation of subcuhuring. The cell morphology and immunocytochemistry tests confirmed that the cultural ceils belonged to epithelial cells. The results indi- cate that the in vitro methods for isolation and culture of tureen epithelial cells in Chinese reindeer are basically established.

关 键 词：中国驯鹿 瘤胃 上皮细胞 原代培养 体外培养 

分 类 号：S825[农业科学—畜牧学]