中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 肉用杂交犊牛小肠上皮细胞的分离培养和鉴定 动物营养学报 2019 (5) 2278-2286
Position: Home > Articles > 肉用杂交犊牛小肠上皮细胞的分离培养和鉴定 Chinese Journal of Animal Nutrition 2019 (5) 2278-2286

肉用杂交犊牛小肠上皮细胞的分离培养和鉴定

作  者：
刘倚帆;谭秀文;游伟;张相伦;姜富贵;宋恩亮;万发春
单  位：
山东省农业科学院畜牧兽医研究所
关键词：
犊牛;小肠上皮细胞;联合消化法;细胞鉴定;核型分析
摘  要：
本试验旨在建立肉用杂交犊牛小肠上皮细胞分离培养和鉴定的方法,为研究犊牛小肠上皮细胞营养吸收、免疫调控及肠道屏障功能提供原代细胞培养模型.选用新生未吮乳的肉用杂交犊牛的空肠组织,应用胶原酶Ⅰ和中性酶联合消化法对小肠上皮细胞进行分离,应用相差消化法和相差贴壁法对犊牛小肠上皮细胞进行纯化.通过细胞形态学观察、生长曲线、免疫荧光及染色体核型分析等方法来鉴定细胞.结果表明:1)应用胶原酶Ⅰ和中性酶联合消化法得到的犊牛小肠上皮细胞生长状况良好,经纯化后得到90%以上纯度的犊牛小肠上皮细胞,并稳定传代至10代以上; 2)犊牛小肠上皮细胞的生长曲线为"S"形,符合细胞增殖规律; 3)免疫荧光鉴定犊牛小肠上皮细胞特异性表达角蛋白13和绒毛蛋白; 4)透射电镜观察发现犊牛小肠上皮细胞边缘有微绒毛结构,细胞与细胞之间的紧密连接结构清晰可见; 5)染色体核型分析显示细胞内含有60条染色体,形态正常,呈二倍体核型.综上所述,应用胶原酶Ⅰ和中性酶联合消化及相差贴壁纯化成功得到犊牛小肠上皮细胞,为研究犊牛小肠上皮细胞营养物质吸收、免疫调控和肠道屏障功能提供了细胞素材.
作  者：
LIU Yifan;TAN Xiuwen;YOU Wei;ZHANG Xianglun;JIANG Fugui;SONG Enliang;WAN Fachun;Institute of Animal Science and Veterinary Medicine,Shandong Academy of Agricultural Sciences;Shandong Key Lab of Animal Disease Control and Breeding;Shandong Provincial Testing Center of Beef Cattle Performance;Shandong Provincial Engineering Technology Center of Animal Healthy Breeding;
关键词：
calf;;intestinal epithelial cell;;combined digestion method;;cell identification;;karyotype analysis
摘  要：
This experiment was designed to establish a method for isolation,culture and identification of hybrid calf intestinal epithelial cells,in order to provide primary cell culture models for the study of intestinal nutrient absorption,immune regulation and intestinal barrier function. Jejunum tissue of newborn hybrid calf was cut down and digested with mixture of collagenase Ⅰ and dispase. The calf intestinal epithelial cells were purified by differential digestion and differential attachment methods. The methods of cell morphology observation,growth curve,immunofluorescence and karyotype analysis were used to identify the cells. The results showed as follows: 1) calf intestinal epithelial cells obtained using mixture of collagenase Ⅰ and dispase digestive method grown well. The calf intestinal epithelial cells could be purified to more than 90%,and could pass more than 10 generations stably. 2) The"S"shape of growth curve of calf intestinal epithelial cells was accordance with the pattern of cell proliferation. 3) The calf intestinal epithelial cells were stained positive for cytokeratin-13 and villin by immunofluorescence identification. 4) Morphological features such as microvilli structure and tight junctions between cells were observed under transmission electron microscope. 5) Karyotype analysis showed that the number of chromosomes with normal morphology and the karyotype was diploid. The above results show that calf intestinal epithelial cells has been successfully obtained by collagenase Ⅰ and dispase combined digestion and differential attachment purification,which can used for the study of intestinal nutrient absorption,immune regulation and the intestinal barrier function.[Chinese Journal of Animal Nutrition,2019,31( 5) : 2278-2286]

相似文章

导出引用
计量
文章访问数: 5
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46