Intestinal epithelial cells (IECs) not only have an absorption function but also act as a physical barrier between the body and the intestinal bacterial flora. Damage to IECs leads to the breakdown of this barrier and has negative effects on animal health. Intestinal epithelial damage is frequently associated with long-term acute stress, such as increased temperature and new stress management models. The intestinal epithelial damage caused by environmental stress has been linked to oxidative stress. Until now, the effects of intestinal epithelial antioxidant activity from feed additives and treatments could be tested in ducks only in vivo because of the lack of in vitro cell culture systems. In this study, we describe our protocol for the easy isolation and culture of IECs from the small intestine of duck embryos. Immunofluorescence was used for the cytological identification of IECs. In addition, IEC marker genes (IAP and CDH1) could also be detected in cultured cells. And cell status assessments were performed, and cell proliferation viability was analyzed by CCK-8 assay. Furthermore, we constructed an oxidative stress model to be used to research the oxidative stress response mechanism, and drugs acting on the cell signal transduction pathway. In conclusion, we have developed an effective and rapid protocol for obtaining duck primary IECs and constructed an oxidative stress model. These IECs exhibit features consistent with epithelial cells and could be used to explore the physiological mechanisms of oxidative stress ex vivo.

中文翻译：

---

鸭肠道上皮细胞的分离，培养，鉴定及氧化应激模型的建立。

肠上皮细胞（IEC）不仅具有吸收功能，而且还充当人体与肠细菌菌群之间的物理屏障。IEC的损坏导致该障碍的破坏，并对动物健康产生负面影响。肠上皮损伤通常与长期急性应激有关，例如体温升高和新的应激管理模型。由环境压力引起的肠上皮损伤与氧化应激有关。到目前为止，由于缺乏体外细胞培养系统，只能在体内对饲料添加剂和处理方法对肠上皮抗氧化活性的影响进行测试。在这项研究中，我们描述了从鸭胚小肠中容易分离和培养IEC的协议。免疫荧光用于IEC的细胞学鉴定。另外，还可以在培养的细胞中检测IEC标记基因（IAP和CDH1）。并进行细胞状态评估，并通过CCK-8分析法分析细胞增殖活力。此外，我们构建了氧化应激模型，用于研究氧化应激反应机制以及作用于细胞信号转导途径的药物。总之，我们已经开发出一种有效且快速的协议来获得鸭的原始IEC，并构建了氧化应激模型。这些IEC具有与上皮细胞一致的特征，可用于探索离体氧化应激的生理机制。并进行细胞状态评估，并通过CCK-8测定法分析细胞增殖活力。此外，我们构建了氧化应激模型，用于研究氧化应激反应机制以及作用于细胞信号转导途径的药物。总之，我们已经开发出一种有效且快速的协议来获得鸭的原始IEC，并构建了氧化应激模型。这些IEC具有与上皮细胞一致的特征，可用于探索离体氧化应激的生理机制。并进行细胞状态评估，并通过CCK-8测定法分析细胞增殖活力。此外，我们构建了氧化应激模型，用于研究氧化应激反应机制以及作用于细胞信号转导途径的药物。总之，我们已经开发出一种有效且快速的协议来获得鸭的原始IEC，并构建了氧化应激模型。这些IEC具有与上皮细胞一致的特征，可用于探索离体氧化应激的生理机制。我们已经开发出一种有效且快速的协议来获得鸭的原始IEC，并构建了氧化应激模型。这些IEC具有与上皮细胞一致的特征，可用于探索离体氧化应激的生理机制。我们已经开发出一种有效且快速的协议来获得鸭的原始IEC，并构建了氧化应激模型。这些IEC具有与上皮细胞一致的特征，可用于探索离体氧化应激的生理机制。