title: Establishment of primary bovine intestinal epithelial cell culture and clone method.
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.
authors: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People's Republic of China.
authors: Kang
authors: K
authors: Zhan
authors: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People's Republic of China.
authors: Miao
authors: M
authors: Lin
authors: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People's Republic of China.
authors: Ming-Mei
authors: MM
authors: Liu
authors: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People's Republic of China.
authors: Yang-Nan
authors: YN
authors: Sui
authors: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People's Republic of China. gqzhao@yzu.edu.
authors: Guo-Qi
authors: GQ
authors: Zhao
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D049109:Cell Proliferation; D048430:Cell Shape; D002478:Cells, Cultured; D002999:Clone Cells; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D007422:Intestines / Q000166:cytology*; D061251:Primary Cell Culture / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction
keywords: Bovine intestinal epithelial cells; Clone; Cytokeratin 18; Model