摘要
采用灌肠消化、组织块消化、机械刮取+胰蛋白酶消化和机械刮取+胶原酶消化等4种方法对仔猪小肠黏膜上皮细胞(Intestinal epithelial cell,IEC)的体外分离培养进行了比较.结果表明,使用机械刮取+胶原酶消化法分离IEC,可简便快速地获得数量大、活性高的小肠上皮细胞,其中原代培养细胞24 h出现贴壁,7～8 d明显增殖,10～12 d贴壁细胞汇合成片,18～21 d细胞呈"铺路石"状;传代培养细胞3 h出现贴壁,3～4 d快速增殖.采用碱性磷酸酶显色反应检测,90%以上的贴壁细胞呈阳性反应.
In order to obtain pure,viable,and functional intestinal epithelial cells（IEC） from piglets,four dissociation methods including coloclysis digestion,organization block digestion,mechanical scrape plus trypsin digestionand and mechanical scrape plus collagenase digestion were tested.The results showed that mechanical scrape plus collagenase digestion was the best method to isolate IEC.About 24 hours after inoculation,IEC attached to the surface of the container.The cells proliferated obviously at 7~8 days of culture,reached 80% of confluence at 10~12 days and showed ＂pavement＂ typical character at 18~21 days.About 3 hours after split from the primarily cultured cells,IEC began to attached,and remarkably proliferated at 3~4 days.Histochemical staining for alkaline phosphatase（AKP） revealed that AKP activity was positive in more than 90% of the attached cells.
出处
《仲恺农业工程学院学报》
CAS
2010年第1期30-33,共4页
Journal of Zhongkai University of Agriculture and Engineering
基金
国家自然科学基金委-广东联合基金(U0731004)资助项目
关键词
猪
小肠
上皮细胞
原代培养
传代培养
pig
small intestine
epithelial cell
primary culture
passage culture
