An ex vivo acid injury and repair (AIR) model using precision-cut lung slices to understand lung injury and repair
File(s)
Accepted version
Author(s)
Kim, Sally
Mongey, Roisin
Griffiths, Mark
Hind, Matthew
Dean, Charlotte
Type
Journal Article
Abstract
Recent advances in cell culture models like air‒liquid interface culture and ex vivo models such as organoids have advanced studies of lung biology; however, gaps exist between these models and tools that represent the complexity of the three‐dimensional environment of the lung. Precision‐cut lung slices (PCLS) mimic the in vivo environment and bridge the gap between in vitro and in vivo models. We have established the acid injury and repair (AIR) model where a spatially restricted area of tissue is injured using drops of HCl combined with Pluronic gel. Injury and repair are assessed by immunofluorescence using robust markers, including Ki67 for cell proliferation and prosurfactant protein C for alveolar type 2/progenitor cells. Importantly, the AIR model enables the study of injury and repair in mouse lung tissue without the need for an initial in vivo injury, and the results are highly reproducible. Here, we present detailed protocols for the generation of PCLS and the AIR model. We also describe methods to analyze and quantify injury in AIR‐PCLS by immunostaining with established early repair markers and fluorescence imaging. This novel ex vivo model is a versatile tool for studying lung cell biology in acute lung injury and for semi‐high‐throughput screening of potential therapeutics. © 2020 Wiley Periodicals LLC.
Date Issued
2020-12-01
Date Acceptance
2020-10-22
Citation
Current protocols in mouse biology, 2020, 10 (4), pp.e85-e85
ISSN
2161-2617
Publisher
Wiley-Blackwell
Start Page
e85
End Page
e85
Journal / Book Title
Current protocols in mouse biology
Volume
10
Issue
4
Copyright Statement
© 2020 Wiley Periodicals LLC. This is the peer reviewed version of the following article, which has been published in final form at https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpmo.85. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.
Sponsor
Royal Brompton & Harefield NHS Foundation Trust
Royal Brompton & Harefield Hospitals Charity
Grant Number
B1064
B1064
Subjects
acid injury
ex vivo model
lung injury
lung tissue repair
precision-cut lung slices
Publication Status
Published
Date Publish Online
2020-11-20