Abstract：
Advancing knowledge of gastrointestinal physiology and its diseases critically depends on the development of precise, species-specific in vitro models that faithfully mimic in vivo intestinal tissues. This is particularly vital for investigating host-pathogen interactions in bovines, which are significant reservoirs for pathogens that pose serious public health risks. Traditional 3D organoids offer limited access to the intestinal epithelium\'s apical surface, a hurdle overcome by the advent of 2D monolayer cultures. These cultures, derived from organoid cells, provide an exposed luminal surface for more accessible study. In this research, a detailed protocol is introduced for creating and sustaining 2D monolayer cultures from cells of bovine small and large intestinal organoids. This method includes protocols for assessing membrane integrity through transepithelial electrical resistance and paracellular permeability alongside immunocytochemistry staining techniques. These protocols lay the groundwork for establishing and characterizing a 2D bovine monolayer culture system, pushing the boundaries of these method applications in biomedical and translational research of public health importance. Employing this innovative approach enables the development of physiologically pertinent in vitro models for exploring both normal and diseased states of cattle intestinal physiology. The implications for biomedical and agricultural advancements are profound, paving the way for more effective treatments for intestinal ailments in cattle, thereby enhancing both animal welfare and food safety.
摘要：
提高胃肠生理学及其疾病的知识关键取决于精确的发展，物种特异性体外模型，忠实地模拟体内肠组织。这对于研究牛的宿主-病原体相互作用尤其重要，这是构成严重公共卫生风险的病原体的重要储库。传统的3D类器官提供有限的进入肠上皮的顶端表面，2D单层文化的出现克服了一个障碍。这些文化,来源于类器官细胞，提供一个暴露的管腔表面更容易接近的研究。在这项研究中,介绍了从牛小肠和大肠类器官的细胞创建和维持2D单层培养物的详细方案。该方法包括通过跨上皮电阻和细胞旁通透性以及免疫细胞化学染色技术评估膜完整性的方案。这些协议为建立和表征2D牛单层培养系统奠定了基础，推动这些方法在生物医学和转化研究中的应用界限。采用这种创新方法可以开发生理相关的体外模型，以探索牛肠道生理的正常和患病状态。对生物医学和农业进步的影响是深远的，为更有效地治疗牛的肠道疾病铺平道路，从而提高动物福利和食品安全。
