A human PCLS model of lung injury and repair for discovery and pharmaceutical research
File(s)
Published version
Author(s)
Bankole, Esther
Wong, Chun Wai
Kim, Sally
Hind, Matthew
Dean, Charlotte
Type
Journal Article
Abstract
Background: The Acid Injury and Repair (AIR) model is an ex vivo model of lung injury and repair, that was previously established using mouse precision-cut lung slices (PCLS). The AIR model provides a bridge between the current in vitro and in vivo models to study the effects of lung injury in 3D lung tissue slices. Here, we show that the AIR model can be adapted for use in human tissue as a translational model for discovery research and drug screening.
Methods: To generate PCLS, resected human lung tissue was coated with alginate hydrogel to form an artificial pleura. Lung tissue was inflated by point injecting 3% agarose, followed by generation of 450-500 μM thick slices of tissue. An isolated area of each slice was injured by brief application of 0.1 M Hydrochloric acid. AIR-PCLS were then washed and cultured for 48 hr before immunostaining to assess proliferating cells (Ki67) alveolar type II/progenitor cell markers (HTII, proSP-C), Lipofibroblasts (ADRP) and endothelial cells (ERG). Viability of PCLS was assessed by both MTT assay and Live/Dead staining.
Results: We show that levels of proliferation do not change in response to acid injury. However, there is a significant increase in the percentage of proSP-C and HTII positive cells in the injured regions of AIR-PCLS. We also identify non-epithelial cell populations; lipofibroblasts and endothelial cells in human AIR-PCLS, to demonstrate that other repair relevant cell types can be identified and tracked in the human AIR (hAIR model).
Conclusions: The hAIR model is an effective ex-vivo tool to study early mechanisms of lung repair following injury. By establishing an area of injured tissue adjacent to uninjured tissue, this model mimics the heterogenous pattern of lung injury frequently present in lung diseases. The hAIR model will facilitate mechanistic studies of human lung repair and provides a valuable pre-clinical model for drug testing.
Methods: To generate PCLS, resected human lung tissue was coated with alginate hydrogel to form an artificial pleura. Lung tissue was inflated by point injecting 3% agarose, followed by generation of 450-500 μM thick slices of tissue. An isolated area of each slice was injured by brief application of 0.1 M Hydrochloric acid. AIR-PCLS were then washed and cultured for 48 hr before immunostaining to assess proliferating cells (Ki67) alveolar type II/progenitor cell markers (HTII, proSP-C), Lipofibroblasts (ADRP) and endothelial cells (ERG). Viability of PCLS was assessed by both MTT assay and Live/Dead staining.
Results: We show that levels of proliferation do not change in response to acid injury. However, there is a significant increase in the percentage of proSP-C and HTII positive cells in the injured regions of AIR-PCLS. We also identify non-epithelial cell populations; lipofibroblasts and endothelial cells in human AIR-PCLS, to demonstrate that other repair relevant cell types can be identified and tracked in the human AIR (hAIR model).
Conclusions: The hAIR model is an effective ex-vivo tool to study early mechanisms of lung repair following injury. By establishing an area of injured tissue adjacent to uninjured tissue, this model mimics the heterogenous pattern of lung injury frequently present in lung diseases. The hAIR model will facilitate mechanistic studies of human lung repair and provides a valuable pre-clinical model for drug testing.
Date Issued
2025-07-05
Date Acceptance
2025-06-23
Citation
Respiratory Research, 2025, 26
ISSN
1465-9921
Publisher
BMC
Journal / Book Title
Respiratory Research
Volume
26
Copyright Statement
© The Author(s) 2025. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
License URL
Publication Status
Published
Article Number
ARTN 237
Date Publish Online
2025-07-01