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

Pediatr Res. 2020 Feb;87(3):511-517. doi: 10.1038/s41390-019-0340-9. Epub 2019 Feb 18.


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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Age Factors
  • Cell Differentiation
  • Cell Lineage
  • Cell Proliferation
  • Cell Separation*
  • Cells, Cultured
  • Epithelial Cells / metabolism
  • Epithelial Cells / physiology*
  • Epithelial Cells / virology
  • Forkhead Transcription Factors / genetics
  • Forkhead Transcription Factors / metabolism
  • Gene Expression Regulation
  • Host-Pathogen Interactions
  • Humans
  • Influenza A Virus, H1N1 Subtype / pathogenicity
  • Influenza, Human / genetics
  • Influenza, Human / metabolism
  • Influenza, Human / virology
  • Keratin-5 / genetics
  • Keratin-5 / metabolism
  • Lung / cytology*
  • Mucin-5B / genetics
  • Mucin-5B / metabolism
  • Phenotype
  • Primary Cell Culture
  • Pulmonary Surfactant-Associated Protein B / genetics
  • Pulmonary Surfactant-Associated Protein B / metabolism
  • RNA-Seq
  • Single-Cell Analysis
  • Tissue Donors*


  • FOXJ1 protein, human
  • Forkhead Transcription Factors
  • KRT5 protein, human
  • Keratin-5
  • MUC5B protein, human
  • Mucin-5B
  • Pulmonary Surfactant-Associated Protein B