Abstract
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.
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This work was partly supported by the National Natural Science Foundation of China (nos. 31301997 and 31572430). Animal slaughter followed the Ethics Committee of the Institute of Yangzhou University.
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Editor: Tetsuji Okamoto
Kang Zhan and Miao Lin contributed equally to this work.
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Zhan, K., Lin, M., Liu, MM. et al. Establishment of primary bovine intestinal epithelial cell culture and clone method. In Vitro Cell.Dev.Biol.-Animal 53, 54–57 (2017). https://doi.org/10.1007/s11626-016-0082-5
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DOI: https://doi.org/10.1007/s11626-016-0082-5