biostudies-literatureKim JNational Research Foundation of KoreaSeoul National University Bundang Hospital research fund1991https://www.ebi.ac.uk/biostudies/studies/S-EPMC11639892biostudies-literatureUnknown13(23)To develop in vitro respiratory models, it is crucial to identify the factors involved in epithelial cell differentiation. In this study, we comprehensively analyzed the effects of air-liquid interface (ALI) culture on epithelial cell differentiation using single-cell RNA sequencing (scRNA-seq). ALI culture induced a pronounced shift in cell composition, marked by a fivefold increase in ciliated cells and a reduction of more than half in basal cells. Transcriptional signatures associated with epithelial cell differentiation, analyzed using iPathwayGuide software, revealed the downregulation of <i>VEGFA</i> and upregulation of <i>CDKN1A</i> as key signals for epithelial differentiation. Our findings highlight the efficacy of the ALI culture for replicating the human lung airway epithelium aCellsTranscriptomic Analysis of Air-Liquid Interface Culture in Human Lung Organoids Reveals Regulators of Epithelial Differentiation.PMC1163989202-2023-0049No. 02-2023-00492021K1A4A7A02097757Kim BKoo BKEo EYKim HJCho YJLee HKim JCho SfalseTranscriptomic Analysis of Air-Liquid Interface Culture in Human Lung Organoids Reveals Regulators of Epithelial Differentiation.To develop in vitro respiratory models, it is crucial to identify the factors involved in epithelial cell differentiation. In this study, we comprehensively analyzed the effects of air-liquid interface (ALI) culture on epithelial cell differentiation using single-cell RNA sequencing (scRNA-seq). ALI culture induced a pronounced shift in cell composition, marked by a fivefold increase in ciliated cells and a reduction of more than half in basal cells. Transcriptional signatures associated with epithelial cell differentiation, analyzed using iPathwayGuide software, revealed the downregulation of <i>VEGFA</i> and upregulation of <i>CDKN1A</i> as key signals for epithelial differentiation. Our findings highlight the efficacy of the ALI culture for replicating the human lung airway epithelium a2024-01-01T00:00:00Z2024 Dec2026-07-09T10:29:40.911Z2025-04-04T01:26:11.206ZS-EPMC116398923968273910.3390/cells13231991