| 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。 |
| 关键词: 山羊 小肠上皮细胞 细胞角蛋白18 波形蛋白 肌间线蛋白 细胞模型 |
| DOI：10.11841/j.issn.1007-4333.2017.10.10 |
| 投稿时间：2016-09-01 |
| 基金项目:江苏高校品牌专业建设工程资助项目（PPZY2015A083） |
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| Isolation culture and identification of goat intestinal epithelial cells |
|
LI Xiaofen1, ZHAN Kang2, ZHANG Xiangying1, TAO yong1, YANG Xiaozhi1, GU Wenjie1
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| (1.Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, China;2.College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China) |
| Abstract: |
| The aim of this study was to establish a primary goat intestinal epithelial cell culture model in vitro and investigate goat intestinal nutrient absorption,immune regulation and relationship between enteric microorganism and intestinal epithelial cells.Intestinal tissue pieces were used to obtain goat intestinal epithelial cells in vitro.The cells isolated were further separated and purified by single clone method.These cells isolated were then identified by cell morphology,cytokeratin 18 immunofluorescence,vimentin,desmin and cell proliferation curve.The results showed that goat intestinal epithelial cells was successfully cultured and passaged generations 10 by tissue adherent method;No expressions of vimentin and desmin genes in goat intestinal epithelial cells were detected using RT-PCR;Positive reaction of goat intestinal epithelial cells against cytokeratine 18 antigen,green fluorescent under fluorescence microscope was discovered.The eighth generation cells still displayed characteristics of epithelial cells.To the tenth goat intestinal epithelial cells cytoplasm became bigger and started to senesce.In conclusions,the cells isolated were successfully passaged generation 10 by using the tissue adherent method,which provided cell model in vitro for understanding the mechanisms of nutrient substance absorption and regulation in goat intestinal epithelial cells. |
| Key words: goat intestinal epithelial cell cytokeratin 18 vimentin desmin cell model |
