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Pediatr Res. 2019 Feb 18. doi: 10.1038/s41390-019-0340-9. [Epub ahead of print]

A novel in vitro model of primary human pediatric lung epithelial cells.

Author information

1
Division of Neonatology, University of Rochester Medical Center, Rochester, NY, USA.
2
Program in Pediatric Molecular and Personalized Medicine, Department of Pediatrics, University of Rochester Medical Center, Rochester, NY, USA.
3
Center for Oral Biology, University of Rochester Medical Center, Rochester, NY, USA.
4
Department of Biomedical Genetics, University of Rochester Medical Center, Rochester, NY, USA.
5
Center for Pediatric Biomedical Research, University of Rochester Medical Center, Rochester, NY, USA.
6
UR Genomics Research Center, University of Rochester Medical Center, Rochester, NY, USA.
7
Division of Neonatology, University of Rochester Medical Center, Rochester, NY, USA. Tom_Mariani@urmc.rochester.edu.
8
Program in Pediatric Molecular and Personalized Medicine, Department of Pediatrics, University of Rochester Medical Center, Rochester, NY, USA. Tom_Mariani@urmc.rochester.edu.
9
Center for Pediatric Biomedical Research, University of Rochester Medical Center, Rochester, NY, USA. Tom_Mariani@urmc.rochester.edu.

Abstract

BACKGROUND:

Current in vitro human lung epithelial cell models derived from adult tissues may not accurately represent all attributes that define homeostatic and disease mechanisms relevant to the pediatric lung.

METHODS:

We report methods for growing and differentiating primary Pediatric Human Lung Epithelial (PHLE) cells from organ donor infant lung tissues. We use immunohistochemistry, flow cytometry, quantitative RT-PCR, and single cell RNA sequencing (scRNAseq) analysis to characterize the cellular and transcriptional heterogeneity of PHLE cells.

RESULTS:

PHLE cells can be expanded in culture up to passage 6, with a doubling time of ~4 days, and retain attributes of highly enriched epithelial cells. PHLE cells can form resistant monolayers, and undergo differentiation when placed at air-liquid interface. When grown at Air-Liquid Interface (ALI), PHLE cells expressed markers of airway epithelial cell lineages. scRNAseq suggests the cultures contained 4 main sub-phenotypes defined by expression of FOXJ1, KRT5, MUC5B, and SFTPB. These cells are available to the research community through the Developing Lung Molecular Atlas Program Human Tissue Core.

CONCLUSION:

Our data demonstrate that PHLE cells provide a novel in vitro human cell model that represents the pediatric airway epithelium, which can be used to study perinatal developmental and pediatric disease mechanisms.

PMID:
30776794
DOI:
10.1038/s41390-019-0340-9

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