Platform Effects on Regeneration by Pulmonary Basal Cells as Evaluated by Single-Cell RNA Sequencing.

Abstract

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.

• Authors: Allison M Greaney, Taylor S Adams, Micha Sam Brickman Raredon, Elise Gubbins, Jonas C Schupp, Alexander J Engler, Mahboobe Ghaedi, Yifan Yuan, Naftali Kaminski, Laura E Niklason
• Journal: Cell reports (Cell Rep) Vol. 30 Issue 12 Pg. 4250-4265.e6 (03 24 2020) ISSN: 2211-1247 [Electronic] United States
• PMID: 32209482 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.

Topics

• Animals
• Cell Differentiation
• Epithelial Cells (cytology, metabolism)
• Epithelium (physiology)
• Lung (cytology)
• Male
• RNA-Seq
• Rats
• Rats, Sprague-Dawley
• Regeneration
• Single-Cell Analysis
• Tissue Engineering
• Trachea (cytology)
• Transcriptome (genetics)