Enterocytes exert an absorptive and protective function in the intestine, and they encounter many different challenging factors such as feed, bacteria, and parasites. An intestinal epithelial in vitro model can help to understand how enterocytes are affected by these factors and contribute to the development of strategies against pathogens. The present study describes a novel method to culture and maintain primary chicken enterocytes and their characterization by immunofluorescence and biomolecular approaches. Starting from 19-day-old chicken embryos it was possible to isolate viable intestinal cell aggregates that can expand and produce a self-maintaining intestinal epithelial cell population that survives until 12 days in culture. These cells resulted positive in immunofluorescence to Cytokeratin 18, Zonula occludens 1, Villin, and Occludin that are common intestinal epithelial markers, and negative to Vimentin that is expressed by endothelial cells. Cells were cultured also on Transwell® permeable supports and trans-epithelial electrical resistance, was measured. This value gradually increased reaching 64 Ω*cm2 7 days after seeding and it remained stable until day 12. Based on these results it was confirmed that it is possible to isolate and maintain chicken intestinal epithelial cells in culture and that they can be suitable as in vitro intestinal model for further studies.

中文翻译：

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鸡肠上皮细胞的分离、培养和表征

肠上皮细胞在肠道中发挥吸收和保护功能，它们会遇到许多不同的挑战因素，例如饲料、细菌和寄生虫。肠上皮体外模型有助于了解肠细胞如何受到这些因素的影响，并有助于制定针对病原体的策略。本研究描述了一种培养和维持原代鸡肠细胞的新方法，并通过免疫荧光和生物分子方法对其进行表征。从 19 天大的鸡胚胎开始，可以分离出活的肠道细胞聚集体，这些细胞聚集体可以扩张并产生自我维持的肠道上皮细胞群，在培养中存活 12 天。这些细胞对细胞角蛋白 18、封闭带 1、绒毛蛋白、和 Occludin 是常见的肠上皮标志物，对内皮细胞表达的波形蛋白呈阴性。细胞也在 Transwell® 渗透性支持物上培养并测量跨上皮电阻。该值在接种 7 天后逐渐增加，达到 64 Ω*cm2 并保持稳定直到第 12 天。基于这些结果，证实可以分离和维持培养中的鸡肠上皮细胞，并且它们可以适用于用于进一步研究的体外肠道模型。