新疆农业科学2012，Vol.49Issue(2)：318-323,6.DOI:CNKI:65-1097/S.20120302.2238.011
新疆军垦细毛羊肾小管上皮细胞的原代培养及鉴定
Primary Culture and Identification of Renal Tubular Epithelial Cells of Xinjiang Fine -wool Sheep
摘要
Abstract
[Objective] The projected aimed to explore an effective and practical method for primary culture of Xinjiang fine - wool sheep renal tubular epithelial cells, so as to establish the basis for the study of diseases caused by kidney damage. [ Method] Renal cortex of fine - wool sheep at 5 - 8 months old were chosen and cut into pieces of 1 - 2 mm3 Traditional and modified tissue culture methods were used, and cultured in DMEM supplemented with 15% -20%newborn calf serum, without grinding or using enzymes and chemicals. All this was done based on the characteristics of epithelial cell. The renal tubular epithelial cells were separated and purified by combining scraping with the different time for cell digestion and attachment, and then filtered by 0.037 mm aperture sieve to remove glomeruli epithelial cells. The isolated cells were identified by immunohistochemistry. [Result]Compared with the traditional one, the modified method could lead to this result : cells adhered early and grew fast, low pollution was produced and high efficiency of cell culture could be reached. Renal tubular epithelial cells were successfully cultivated and isolated, and the purity reached as high as 90%. The cultured cells displayed polygon, cobblestone layout and strong refraction, tight connection, and they were successively propagated nine times. They were identified as cells of epithelial origin by immunohistochemistry(CK -18). [Conclusion] Compared with traditional tissue culture and enzymatic digestion culture, this economic, simple and pollution - free culture system is an effective one to cultivate renal tubular epithelial cells with high activity and purity.关键词
肾小管上皮细胞/原代培养/分离纯化/细毛羊Key words
renal tubular epithelial cell/ primary culture/ isolation and purification/ fine - wool sheep分类
农业科技引用本文复制引用
潘奇,李宏建,潘晓亮,张书信,崔艳霞,王振国,徐亚楠..新疆军垦细毛羊肾小管上皮细胞的原代培养及鉴定[J].新疆农业科学,2012,49(2):318-323,6.基金项目
新疆兵团科技局重点项目(9808010124) （9808010124）
