A comparative study of in vitro air-liquid interface culture models of the human airway epithelium evaluating cellular heterogeneity and gene expression at single cell resolution.
A comparative study of in vitro air-liquid interface culture models of the human airway epithelium evaluating cellular heterogeneity and gene expression at single cell resolution.
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人气道上皮体外气液界面培养模型的比较研究，以单细胞分辨率评估细胞异质性和基因表达。
DOI：
10.1101/2023.02.27.530299
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Rosenberg,BradR
中科院分区：
文献类型：
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作者：
Prescott,RachelA;Pankow,AlecP;deVries,Maren;Crosse,Keaton;Patel,RoosheelS;Alu,Mark;Loomis,Cynthia;Torres,Victor;Koralov,Sergei;Ivanova,Ellie;Dittmann,Meike;Rosenberg,BradR
气道上皮由不同类型的细胞组成，具有调节体内平衡和保护呼吸道病原体的专门功能。人气道上皮细胞培养物在气液界面（HAE）是一种生理相关的体外模型，这种异质组织，使许多研究的气道疾病1 - 7。HAE培养物通常来源于原代上皮细胞，其相对有限的传代能力可能限制实验方法和研究设计。BCi-NS1。1，一种先前描述并广泛使用的工程化以表达hTERT的基础细胞系，表现出延长的传代寿命，同时保留分化为HAE 5的能力。然而，HAE中的基因表达和先天免疫功能源自BCi-NS1。1与原代细胞的比较尚未完全表征。在这里，结合单细胞RNA-Seq（scRNA-Seq），免疫组织化学和功能实验，我们以高分辨率证实了BCi-NS1。1和原代HAE培养物在形态学、细胞类型组成和总体转录模式方面很大程度上相似。同时我们观察到BCi-NS1中几个干扰素刺激基因的细胞类型特异性表达差异。1 HAE培养物中，我们没有观察到对甲型流感病毒和金黄色葡萄球菌感染的易感性的显著差异。综上所述，我们的结果进一步支持BCi-NS1。1-衍生的HAE培养物作为研究气道感染性疾病的有价值的工具。
The airway epithelium is composed of diverse cell types with specialized functions that mediate homeostasis and protect against respiratory pathogens. Human airway epithelial cultures at air-liquid interface (HAE) are a physiologically relevant in vitro model of this heterogeneous tissue, enabling numerous studies of airway disease 1–7. HAE cultures are classically derived from primary epithelial cells, the relatively limited passage capacity of which can limit experimental methods and study designs. BCi-NS1. 1, a previously described and widely used basal cell line engineered to express hTERT, exhibits extended passage lifespan while retaining capacity for differentiation to HAE 5. However, gene expression and innate immune function in HAE derived from BCi-NS1. 1 versus primary cells have not been fully characterized. Here, combining single cell RNA-Seq (scRNA-Seq), immunohistochemistry, and functional experimentation, we confirm at high resolution that BCi-NS1. 1 and primary HAE cultures are largely similar in morphology, cell type composition, and overall transcriptional patterns. While we observed cell-type specific expression differences of several interferon stimulated genes in BCi-NS1. 1 HAE cultures, we did not observe significant differences in susceptibility to infection with influenza A virus and Staphylococcus aureus. Taken together, our results further support BCi-NS1. 1-derived HAE cultures as a valuable tool for the study of airway infectious disease.