奶牛肾小管上皮细胞的原代培养和鉴定  

Primary Culture and Identification of Cow Renal Tubular Epithelial Cells

作　　者：石浪涛[1] 柳东阳[1] 席照房[1] 郭定宗[1] 

机构地区：[1]华中农业大学动物营养代谢病研究中心,武汉430070

出　　处：《中国奶牛》2011年第18期34-37,共4页China Dairy Cattle

摘　　要：为建立奶牛肾小管上皮细胞的原代培养、传代以及鉴定方法,本研究将肾脏皮质剪碎,并网筛过滤选取肾小管后分别进行直接培养、胶原酶1消化培养、胰蛋白酶消化培养和胰蛋白酶和0.02%EDTA混合消化培养,分别应用上述四种方法进行原代培养和传代培养,然后采用CK-18、PCK和Vimentin三种蛋白进行免疫组化鉴定奶牛肾小管上皮细胞。结果表明,采用消化培养方法都能够得到奶牛肾小管上皮细胞,但是胶原酶1消化得到的活细胞更多,能更快地成功培养奶牛肾小管上皮细胞。胶原酶1消化法是培养奶牛肾小管上皮细胞的理想培养法。The objective of this study was to establish a method for the primary culture, subculture and identification of cow renal tubular epithelial cells. Harvested renal tubular segments by mechanical separation, four methods （Directly culture, digest by collagenase 1, digest by trypsin, digest by trypsin ＋ EDTA）were conducted to primary culture the cow renal tubular epithelial cells. By cellular immnnocytochemistry, CK-18, PCK and Vimentin were used to identify the types of cells. Result showed that the three chemical digestion methods can successfully culture cow renal tubular epithelial cells, but collagenase 1 was best. Collagenase 1 digesting was an ideal method for the primary culture of cow renal tubular epithelial cells.

关 键 词：奶牛 肾小管上皮细胞 胶原酶1 CK-18 PCK VIMENTIN 

分 类 号：S858.232.161[农业科学—临床兽医学]