| 鸽小肠上皮细胞的分离培养与鉴定 |
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| 引用本文： | 刘磊,王讯,罗毅,梁继元,颜培祺,李昕怡,刘薇,刘辰恺,李佳佳.鸽小肠上皮细胞的分离培养与鉴定[J].浙江农业学报,2021,33(2):239. |
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| 作者姓名： | 刘磊 王讯 罗毅 梁继元 颜培祺 李昕怡 刘薇 刘辰恺 李佳佳 |
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| 作者单位： | 1.四川农业大学 动物科技学院,四川 成都 6111302.畜禽遗传资源发掘与创新利用四川省重点实验室,四川 成都 611130 |
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| 基金项目： | 四川农业大学双支计划(1921993255) |
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| 摘 要： | 本试验旨在建立鸽小肠上皮细胞体外分离培养和鉴定的方法体系,为研究鸽小肠上皮细胞的营养转运吸收、细胞增殖代谢等机制提供原代细胞模型。采用胶原酶消化法和组织块贴壁法分离鸽小肠上皮细胞,并通过相差消化法对其纯化,最后运用免疫荧光分析和基因表达分析鉴定鸽小肠上皮细胞。结果表明,2种方法均可成功分离培养鸽小肠上皮细胞,细胞呈铺路石状排列;细胞角蛋白8免疫荧光染色呈阳性;紧密连接蛋白基因CLDN-3和CLDN-4均有表达;细胞生长曲线呈“S”形。综上所述,本试验成功建立了鸽小肠上皮细胞体外分离培养体系,为后续开展雏鸽肠道营养研究提供了细胞模型。
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| 关 键 词： | 鸽 小肠上皮细胞 原代培养 细胞鉴定 |
| 收稿时间： | 2020-06-20 |
Isolation,culture and identification of pigeon intestine epithelium cells |
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| LIU Lei,WANG Xun,LUO Yi,LIANG Jiyuan,YAN Peiqi,LI Xinyi,LIU Wei,LIU Chenkai,LI Jiajia.Isolation,culture and identification of pigeon intestine epithelium cells[J].Acta Agriculturae Zhejiangensis,2021,33(2):239. |
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| Authors: | LIU Lei WANG Xun LUO Yi LIANG Jiyuan YAN Peiqi LI Xinyi LIU Wei LIU Chenkai LI Jiajia |
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| Institution: | 1. College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China
2. Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Chengdu 611130, China |
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| Abstract: | To establish a set of method to isolate, culture and identify pigeon intestine epithelium cells, and provide a primary cell model to research the mechanism of nutrient transportation and absorption, cell proliferation and metabolism in vitro, pigeon intestine epithelium cells were isolated by collagenase digestion and tissue explants adherent method. Isolated pigeon intestine epithelium cells were identified by immunofluorescence analysis and PCR analysis. The results showed that pigeon intestine epithelium cells were successfully isolated by using collagenase digestion and tissue explants adherent method. The isolated cells were arranged in the shape of paving stones. Cytokeratin 8 immunofluorescence staining was positive, and both CLDN-3 and CLDN-4 gene of tight junction proteins were expressed. The cell growth curve was S shape. In summary, we successfully established a set of method to isolate, culture and identify pigeon intestine epithelium cells, and provided a cell model for the further research on intestinal nutrition of pigeon squabs. |
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| Keywords: | pigeon intestine epithelium cells primary culture cell identification |
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