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| Primary Culture and Identification of Intestinal Epithelial Cells in Sea Bass Lateolabrax maculatus |
| LI Sha1, LIU Ziyan1, YANG Hongling1,2, CAI Guohe1, NIE Qingjie1, ZHANG Chunxiao1,2, SUN Yunzhang1,2 |
1. Xiamen Key Laboratory of Feed Detection and Safety Evaluation, Fisheries College, Jimei University, Xiamen 361021, China;
2. Key Laboratory of Healthy Mariculture in the East China Sea, Ministry of Agriculture and Rural Affairs, Fisheries College, Jimei University, Xiamen 361021, China |
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Abstract The aim of this study was to establish a stable and reliable primary culture method for intestinal epithelial cells (IECs) of sea bass Lateolabrax maculatus. The intestinal epithelial cells of sea bass were cultured and evaluated by tissue block method and enzyme digestion method, the optimal digestion solution and digestion time of enzyme digestion method were determined, and the obtained cell suspension was cultured in L-15 medium. The cells were identified by morphological observation, transmission electron microscopy and alkaline phosphatase staining methods. The results showed that the unstable and long migration time of the cell migration were observed in tissue block method; the optimal digestion solution was collagenase Ⅰ and Ⅳ combined digestion solution, with the optimal digestion time of 45 min. The cells obtained by collagenase Ⅰ and Ⅳ combined digestion were closely connected and arranged in order, showing the typical "paving stone" shape of epithelial cells, with relatively independent cells and clear boundaries. Transmission electron microscopy and alkaline phosphatase staining further confirmed that the cultured cells were intestinal epithelial cells. In conclusion, the primary intestinal epithelial cells with consistent initial conditions and vigorous growth were obtained after 45 min of digestion with collagenase Ⅰ and Ⅳ digestion solution and following by 48 h culture.
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Received: 05 February 2023
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