山羊小肠上皮细胞分离培养与鉴定

发布时间：2018-02-16 22:24

本文关键词： 山羊小肠上皮细胞细胞角蛋白波形蛋白肌间线蛋白细胞模型　出处：《中国农业大学学报》2017年10期 　论文类型：期刊论文

【摘要】：为研究山羊小肠上皮细胞营养吸收调控及其与肠道微生物之间的关系,提供原代细胞培养模型,采用组织块接种来获得山羊小肠上皮细胞,利用有限稀释法来克隆山羊小肠上皮细胞,并通过细胞形态学以及细胞角蛋白18、波形蛋白、肌间线蛋白和细胞生长曲线来鉴定山羊小肠上皮细胞。结果表明:1)采用组织块接种能够分离纯化得到山羊小肠上皮细胞并稳定传至大约10代;2)RT-PCR检测发现山羊小肠上皮细胞不能够表达波形蛋白和肌间线蛋白;3)在正置显微镜下观察到山羊小肠上皮细胞能够产生细胞角蛋白18绿色荧光。研究发现,培养至第8代的山羊小肠上皮细胞仍然保持着上皮细胞的特征,至10代山羊小肠上皮细胞质变大,细胞几乎无法增殖,细胞开始凋亡。综上所述,采用组织块接种能够获得山羊小肠上皮细胞并正常传至第10代,可为研究山羊小肠上皮细胞营养物质吸收和调控机理提供细胞素材。
[Abstract]:In order to study the regulation of nutrient absorption of goat intestinal epithelial cells and their relationship with intestinal microorganism, the primary culture model of goat small intestinal epithelial cells was obtained by tissue block inoculation. Goat small intestinal epithelial cells were cloned by a limited dilution method, and by cell morphology and cytokeratin 18, vimentin. Myofiline and cell growth curves were used to identify goat small intestinal epithelial cells. The results showed that small intestinal epithelial cells could be isolated and purified by tissue mass inoculation and transferred steadily to about 10 passages of small intestine by RT-PCR. Epithelial cells were not able to express vimentin and myoflinin-3) under a positive microscope, goat small intestinal epithelial cells were observed to produce cytokeratin 18 green fluorescence. Goat small intestinal epithelial cells cultured up to the 8th passage still retain the characteristics of epithelial cells, and by the 10th passage goat intestinal epithelial cells become larger, the cells can hardly proliferate, and the cells begin to apoptosis. Goat small intestinal epithelial cells could be obtained by tissue mass inoculation and passed to the tenth passage, which could provide material for studying the absorption and regulation mechanism of nutrients in goat intestinal epithelial cells.
【作者单位】：江苏农牧科技职业学院;扬州大学动物科学与技术学院;
【基金】：江苏高校品牌专业建设工程资助项目(PPZY2015A083)
【分类号】：S827

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