中文摘要：研究分析了气液界面培养系统中影响绵羊气管上皮细胞生长和增殖的关键因素，结果发现：（1）与上皮细胞增殖和纤毛增厚相关的上皮生长因子具有剂量依赖效应；（2）上皮细胞增殖过程中对视黄酸（RA）的要求极为严格，缺乏视黄酸会导致复层鳞状上皮的形成、纤毛变少；（3）生长基质膜孔密度对细胞增殖的影响也较大，膜孔厚度增加会增加纤毛内嵌卷曲水平、增加细胞层的均匀度。 本研究的结果发现了绵羊气管上皮细胞与其他上皮细胞模型在气液界面培养中培养条件的重要区别，这些结果表明绵羊气管上皮细胞的表型可以通过培养条件的精确调控进行调节，因此可以体外调控定制绵羊气管上皮细胞感染模型。
 
外文摘要：Respiratory tract infections are of significant concern in the agriculture industry. There is a requirement for the development of well-characterised in vitro epithelial cell culture models in order to dissect the diverse molecular interactions occurring at the host-pathogen interface in airway epithelia. We have analysed key factors that influence growth and differentiation of ovine tracheal epithelial cells in an air-liquid interface (ALI) culture system. Cellular differentiation was assessed at 21 days post-ALI, a time-point which we have previously shown to be sufficient for differentiation in standard growth conditions. We identified a dose dependent response to epidermal growth factor (EGF) in terms of both epithelial thickening and ciliation levels. Maximal ciliation levels were observed with 25 ng m(-1) EGF. We identified a strict requirement for retinoic acid (RA) in epithelial differentiation as RA exclusion resulted in the formation of a stratified squamous epithelium, devoid of cilia. The pore-density of the growth substrate also had an influence on differentiation as high pore-density inserts yielded higher levels of ciliation and more uniform cell layers than low pore-density inserts. Differentiation was also improved by culturing the cells in an atmosphere of sub ambient oxygen concentration. We compared two submerged growth media and observed differences in the rate of proliferation/expansion, barrier formation and also in terminal differentiation. Taken together, these results indicate important differences between the response of ovine tracheal epithelial cells and other previously described airway epithelial models, to a variety of environmental conditions. These data also indicate that the phenotype of ovine tracheal epithelial cells can be tailored in vitro by precise modulation of growth conditions, thereby yielding a customisable, potential infection model.
外文关键词：IN-VITRO; MUCOCILIARY DIFFERENTIATION; TRACHEAL EPITHELIUM; FACTOR RECEPTOR; RETINOIC ACID; BIGHORN SHEEP; BASAL-CELLS; STEM-CELLS; CULTURES; TRIIODOTHYRONINE
作者：O'Boyle, N; Sutherland, E; Berry, CC; Davies, RL
作者单位：苏格兰格拉斯哥大学
期刊名称：PLOS ONE
期刊影响因子：2.81
出版年份：2018
出版刊次：3
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