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

Respiratory Research

Research

Maren de Vries¹  Victor Torres¹  Rachel A. Prescott¹  Keaton M. Crosse¹  Meike Dittmann¹  Alec P. Pankow²  Roosheel S. Patel²  Brad R. Rosenberg²  Sergei Koralov³  Cynthia Loomis³  Ellie Ivanova³  Mark Alu³ 

[1] Department of Microbiology, NYU Grossman School of Medicine, 10016, New York, NY, USA;Department of Microbiology, The Icahn School of Medicine at Mount Sinai, 10029, New York, NY, USA;Department of Pathology, NYU Grossman School of Medicine, 10016, New York, NY, USA;

关键词: Airway epithelium;    Innate immunity;    Respiratory infection;    Air–liquid interface epithelial culture;    Single cell transcriptomics;   

DOI  :  10.1186/s12931-023-02514-2

 received in 2023-03-15, accepted in 2023-08-17,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundThe airway epithelium is composed of diverse cell types with specialized functions that mediate homeostasis and protect against respiratory pathogens. Human airway epithelial (HAE) cultures at air–liquid interface are a physiologically relevant in vitro model of this heterogeneous tissue and have enabled numerous studies of airway disease. 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 the capacity for differentiation to HAE. However, gene expression and innate immune function in BCi-NS1.1-derived versus primary-derived HAE cultures have not been fully characterized.MethodsBCi-NS1.1-derived HAE cultures (n = 3 independent differentiations) and primary-derived HAE cultures (n = 3 distinct donors) were characterized by immunofluorescence and single cell RNA-Seq (scRNA-Seq). Innate immune functions were evaluated in response to interferon stimulation and to infection with viral and bacterial respiratory pathogens.ResultsWe confirm at high resolution that BCi-NS1.1- and primary-derived HAE cultures are largely similar in morphology, cell type composition, and overall gene expression patterns. While we observed cell-type specific expression differences of several interferon stimulated genes in BCi-NS1.1-derived HAE cultures, we did not observe significant differences in susceptibility to infection with influenza A virus and Staphylococcus aureus.ConclusionsTaken together, our results further support BCi-NS1.1-derived HAE cultures as a valuable tool for the study of airway infectious disease.

【 授权许可】

CC BY   
© BioMed Central Ltd., part of Springer Nature 2023

【 图 表 】

13750_2023_309_Article_IEq64.gif

Fig. 2

Fig. 4

13570_2023_282_Article_IEq15.gif

Fig. 3

13690_2023_1151_Article_IEq20.gif

13690_2023_1151_Article_IEq9.gif

40798_2023_599_Article_IEq15.gif