Do Airway Epithelium Air-Liquid Cultures Represent the In Vivo Airway Epithelium Transcriptome?
Do Airway Epithelium Air-Liquid Cultures Represent the In Vivo Airway Epithelium Transcriptome?
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DOI：
10.1165/rcmb.2009-0453oc
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发表时间：
2011-04-01
影响因子：
6.4
通讯作者：
Crystal, Ronald G.
中科院分区：
文献类型：
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作者：
Dvorak, Anna;Tilley, Ann E.;Crystal, Ronald G.
体外培养的人气道上皮细胞在气液界面（ALI）形成假复层上皮，形成紧密连接和纤毛，并产生粘蛋白。这些细胞广泛用于分化、损伤和修复的模型。为了评估ALI上皮细胞的转录组与体内气道上皮细胞的转录组的匹配程度，我们使用微阵列将在ALI处培养的人大气道上皮细胞的转录组与通过支气管镜检查和刷拭获得的大气道上皮细胞的转录组进行比较。基因表达谱分析显示，ALI细胞和刷拭细胞的基因表达总体相关性良好，但也存在一定差异。基因表达模式反映了细胞类型比例的差异（ALI具有较高的基底细胞百分比，而刷状细胞具有较高的纤毛细胞百分比），即ALI细胞表达较高水平的基底细胞相关基因，刷状细胞表达较高水平的纤毛相关基因。通路分析显示，ALI细胞的细胞周期和增殖基因的表达增加，而刷状细胞的细胞骨架组织和体液免疫反应基因的表达增加。总体而言，ALI细胞提供了体内气道上皮转录组的良好代表性，但对于一些生物学问题，需要考虑体外和体内环境之间的差异。
Human airway epithelial cells cultured in vitro at the air-liquid interface (ALI) form a pseudostratified epithelium that forms tight junctions and cilia, and produces mucin. These cells are widely used in models of differentiation, injury, and repair. To assess how closely the transcriptome of ALI epithelium matches that of in vivo airway epithelial cells, we used microarrays to compare the transcriptome of human large airway epithelial cells cultured at the ALI with the transcriptome of large airway epithelium obtained via bronchoscopy and brushing. Gene expression profiling showed that global gene expression correlated well between ALI cells and brushed cells, but with some differences. Gene expression patterns mirrored differences in proportions of cell types (ALIs have higher percentages of basal cells, whereas brushed cells have higher percentages of ciliated cells), that is, ALI cells expressed higher levels of basal cell-related genes, and brushed cells expressed higher levels of cilia-related genes. Pathway analysis showed that ALI cells had increased expression of cell cycle and proliferation genes, whereas brushed cells had increased expression of cytoskeletal organization and humoral immune response genes. Overall, ALI cells provide a good representation of the in vivo airway epithelial transcriptome, but for some biologic questions, the differences between in vitro and in vivo environments need to be considered.