Temporal dynamics of ovine airway epithelial cell differentiation at an air-liquid interface.
Temporal dynamics of ovine airway epithelial cell differentiation at an air-liquid interface.
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DOI：
10.1371/journal.pone.0181583
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Davies RL
中科院分区：
文献类型：
--
作者：
O'Boyle N;Sutherland E;Berry CC;Davies RL
呼吸道和肺受到多种病理学的影响，对人类和动物的福利都有广泛的影响。开发和详细表征的细胞培养模型研究这些形式的疾病是至关重要的。近年来，使用气液界面（ALI）培养的气道上皮细胞已显着增加，因为这种培养方法的结果在形成一个高度代表性的，器官型的体外模型系统。在这项研究中，我们已经扩大了以前的知识，分化羊气管上皮细胞的分化过程中分析了一个广泛的时间过程中的ALI。我们观察到假复层上皮纤毛和细胞层厚度的增加，从9天后ALI纤毛接近最大水平，在第24天。在粘液产生方面观察到类似的模式，在第12天出现强染色的PAS阳性细胞。扫描电镜超微结构分析证实，纤毛细胞和粘液球的存在下，在此时间范围内的上皮表面。当细胞层融合时，跨上皮电阻（TEER）达到峰值1049 Ω × cm 2，随后随着分化的进行而降低，并稳定在约200 Ω × cm 2。重要的是，在45天的时间过程中几乎没有观察到退化或去分化，表明该模型适合于长期实验。
The respiratory tract and lungs are subject to diverse pathologies with wide-ranging implications for both human and animal welfare. The development and detailed characterization of cell culture models for studying such forms of disease is of critical importance. In recent years the use of air-liquid interface (ALI)-cultured airway epithelial cells has increased markedly, as this method of culture results in the formation of a highly representative, organotypic in vitro model system. In this study we have expanded on previous knowledge of differentiated ovine tracheal epithelial cells by analysing the progression of differentiation over an extensive time course at an ALI. We observed a pseudo-stratified epithelium with ciliation and a concurrent increase in cell layer thickness from 9 days post-ALI with ciliation approaching a maximum level at day 24. A similar pattern was observed with respect to mucus production with intensely stained PAS-positive cells appearing at day 12. Ultrastructural analysis by SEM confirmed the presence of both ciliated cells and mucus globules on the epithelial surface within this time-frame. Trans-epithelial electrical resistance (TEER) peaked at 1049 Ω × cm2 as the cell layer became confluent, followed by a subsequent reduction as differentiation proceeded and stabilization at ~200 Ω × cm2. Importantly, little deterioration or de-differentiation was observed over the 45 day time-course indicating that the model is suitable for long-term experiments.