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Isolation and Enrichment of Human Lung Epithelial Progenitor Cells for Organoid Culture
Published on: July 21, 2020
A novel in vitro model of primary human pediatric lung epithelial cells
Qian Wang1,2,3, Soumyaroop Bhattacharya1,2, Jared A Mereness1,2,4
1Division of Neonatology, University of Rochester Medical Center, Rochester, NY, USA.
Insights
Researchers developed a new method to grow and differentiate pediatric human lung epithelial (PHLE) cells. These cells accurately model the pediatric airway, aiding research into lung development and disease.
Area of Science:
- Pulmonary Medicine
- Cell Biology
- Developmental Biology
Background:
- Current in vitro models using adult lung cells may not fully represent pediatric lung physiology.
- Understanding pediatric lung development and disease requires accurate cellular models.
Purpose of the Study:
- To establish and characterize primary Pediatric Human Lung Epithelial (PHLE) cells from infant lung tissue.
- To provide a novel in vitro model for studying pediatric lung mechanisms.
Main Methods:
- Primary PHLE cells were isolated from infant lung organ donors.
- Cells were cultured, expanded, and differentiated at air-liquid interface (ALI).
- Characterization involved immunohistochemistry, flow cytometry, RT-PCR, and single-cell RNA sequencing (scRNAseq).
Main Results:
- PHLE cells were successfully expanded and retained epithelial characteristics.
- Differentiated PHLE cells at ALI expressed airway epithelial lineage markers.
- scRNAseq identified four main sub-phenotypes (FOXJ1, KRT5, MUC5B, SFTPB).
Conclusions:
- PHLE cells offer a unique human in vitro model of the pediatric airway epithelium.
- This model is suitable for investigating perinatal development and pediatric lung diseases.
- These cells are accessible to the research community via the Developing Lung Molecular Atlas Program.
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.
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