Cell Reports
Volume 30, Issue 12, 24 March 2020, Pages 4250-4265.e6
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Article
Platform Effects on Regeneration by Pulmonary Basal Cells as Evaluated by Single-Cell RNA Sequencing

https://doi.org/10.1016/j.celrep.2020.03.004Get rights and content
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Highlights

  • peBCs are an expandable plastic cell type to study pulmonary epithelial regeneration

  • scRNA-seq enables high-res comparison of engineered tissues to native benchmarks

  • peBCs in organoid and ALI show evidence of non-physiologic assimilation by scRNA-seq

  • peBCs in engineered lung and trachea yield more physiologic epithelial regeneration

Summary

Cell-based therapies have shown promise for treating myriad chronic pulmonary diseases through direct application of epithelial progenitors or by way of engineered tissue grafts or whole organs. To elucidate environmental effects on epithelial regenerative outcomes in vitro, here, we isolate and culture a population of pharmacologically expanded basal cells (peBCs) from rat tracheas. At peak basal marker expression, we simultaneously split peBCs into four in vitro platforms: organoid, air-liquid interface (ALI), engineered trachea, and engineered lung. Following differentiation, these samples are evaluated using single-cell RNA sequencing (scRNA-seq) and computational pipelines are developed to compare samples both globally and at the population level. A sample of native rat tracheal epithelium is also evaluated by scRNA-seq as a control for engineered epithelium. Overall, this work identifies platform-specific effects that support the use of engineered models to achieve the most physiologic differential outcomes in pulmonary epithelial regenerative applications.

Keywords

Pulmonary Cell Biology
Regenerative Medicine
Tissue Engineering
Primary Epithelial Cells

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