Establishment of primary bovine intestinal epithelial cell culture and clone method
Establishment of primary bovine intestinal epithelial cell culture and clone method
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DOI：
10.1007/s11626-016-0082-5
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
K. Zhan;Miao Lin;Ming-mei Liu;Yang-Nan Sui;Guoqi Zhao
中科院分区：
文献类型：
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作者：
K. Zhan;Miao Lin;Ming-mei Liu;Yang-Nan Sui;Guoqi Zhao
本研究的目的是建立牛肠上皮细胞（BIEC）系，并提供一种新的克隆细胞方法。虽然各种策略的牛细胞培养和克隆技术已经报道，这些方法仍然没有建立。在此，我们成功培养了原代biec，并建立了一种新的克隆细胞方法。结果表明，BIECs在第5代左右可以成功培养传代。在培养的第2天，这些细胞聚集体和簇松散地贴壁。大约4 d后，细胞聚集和聚集开始增殖。biec对细胞角蛋白18、e -钙粘蛋白呈阳性反应，具有上皮样形态特征。此外，脂肪酸结合蛋白（FABPs）、绒毛蛋白（villin）和肠肽酶（IP）带在biec中呈阳性。本研究结果表明，BIEC培养和克隆方法的建立将适用于其他原代细胞的分离和克隆。因此，这些biec可用于研究牛肠道营养物质的吸收与调节、免疫调节以及牛肠道疾病的发病机制，为体外肠道细胞模型的建立提供依据。
The aim of this study was to establish bovine intestinal epithelial cell (BIEC) line and provide a novel clone cell method. Although various strategies of bovine cell culture and clone techniques have been reported, these methods remain not established. Here, we culture successfully primary BIECs and establish a novel clone cell method. Our result showed that BIECs could be successfully cultured and passaged about generation 5. These cellular aggregates and clusters were adherent loosely at day 2 of culture. Cell aggregates and clusters start to proliferate after approximately 4 d. The BIECs showed positive reaction against cytokeratin 18, E-cadherin, and characteristics of epithelial-like morphology. In addition, the fatty acid-binding proteins (FABPs), villin, and intestinal peptidase (IP) band were positive in BIECs. Our results suggest that the establishment of culturing and clone BIEC methods will apply to isolate and clone other primary cells. These BIECs could therefore contribute to the study of bovine intestinal nutrient absorption and regulation, immune regulation, and the pathogenesis of the bovine intestinal disease, which will provide intestinal cell model in vitro.