{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,17]],"date-time":"2026-05-17T06:45:09Z","timestamp":1779000309397,"version":"3.51.4"},"reference-count":23,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2020,11,20]],"date-time":"2020-11-20T00:00:00Z","timestamp":1605830400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["104931\/Z\/14\/Z"],"award-info":[{"award-number":["104931\/Z\/14\/Z"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000268","name":"Biotechnology and Biological Sciences Research Council","doi-asserted-by":"publisher","award":["BB\/L015129\/1"],"award-info":[{"award-number":["BB\/L015129\/1"]}],"id":[{"id":"10.13039\/501100000268","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["currentprotocols.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["CP Mouse Biology"],"published-print":{"date-parts":[[2020,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Recent advances in cell culture models like air\u2012liquid 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\u2010dimensional environment of the lung. Precision\u2010cut 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\u2010PCLS 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\u2010high\u2010throughput screening of potential therapeutics. \u00a9 2020 Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: Generation of precision\u2010cut lung slices<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: The acid injury and repair model<\/jats:p><jats:p><jats:bold>Basic Protocol 3<\/jats:bold>: Analysis of AIR\u2010PCLS: Immunostaining and imaging<\/jats:p>","DOI":"10.1002\/cpmo.85","type":"journal-article","created":{"date-parts":[[2020,11,21]],"date-time":"2020-11-21T06:03:01Z","timestamp":1605938581000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["An Ex Vivo Acid Injury and Repair (AIR) Model Using Precision\u2010Cut Lung Slices to Understand Lung Injury and Repair"],"prefix":"10.1002","volume":"10","author":[{"given":"Sally Yunsun","family":"Kim","sequence":"first","affiliation":[{"name":"National Heart and Lung Institute Imperial College London  London United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R\u00f3is\u00edn","family":"Mongey","sequence":"additional","affiliation":[{"name":"National Heart and Lung Institute Imperial College London  London United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark","family":"Griffiths","sequence":"additional","affiliation":[{"name":"National Heart and Lung Institute Imperial College London  London United Kingdom"},{"name":"Peri\u2010Operative Medicine Department St Bartholomew's Hospital  London United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthew","family":"Hind","sequence":"additional","affiliation":[{"name":"National Heart and Lung Institute Imperial College London  London United Kingdom"},{"name":"National Institute for Health Research (NIHR) Respiratory Biomedical Research Unit at the Royal Brompton &amp; Harefield NHS Foundation Trust and Imperial College  London United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Charlotte H.","family":"Dean","sequence":"additional","affiliation":[{"name":"National Heart and Lung Institute Imperial College London  London United Kingdom"},{"name":"MRC Harwell Institute Harwell Campus  Oxfordshire United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,11,20]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"crossref","first-page":"e3403","DOI":"10.21769\/BioProtoc.3403","article-title":"Time\u2010lapse imaging of alveologenesis in mouse precision\u2010cut lung slices","volume":"9","author":"Akram K. M.","year":"2019","journal-title":"Bio\u2010protocol"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-09067-3"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1165\/rcmb.2019-0276TR"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2020.03.009"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.119.2.187"},{"key":"e_1_2_7_7_1","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1186\/s13567-019-0733-0","article-title":"Precision cut lung slices: A novel versatile tool to examine host\u2010pathogen interaction in the chicken lung","volume":"51","author":"Bryson K. J.","year":"2020","journal-title":"Veterinary Research"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/sctm.19-0433"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI118009"},{"key":"e_1_2_7_10_1","doi-asserted-by":"crossref","first-page":"P245","DOI":"10.1016\/j.omtm.2018.07.010","article-title":"Use of precision\u2010cut lung slices as an ex vivo tool for evaluating viruses and viral vectors for gene and oncolytic therapy","volume":"10","author":"Gerpe M. C. R.","year":"2018","journal-title":"Molecular Therapy Methods & Clinical Development"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1183\/13993003.00876-2018"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.taap.2008.04.003"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.150201"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1213\/00000539-198907000-00017"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2020.120480"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12931-019-1131-x"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1186\/1465-9921-7-111"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2015.01.002"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1183\/09031936.00093911"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1242\/dmm.006213"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12931-016-0358-z"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2016.10.046"},{"key":"e_1_2_7_23_1","doi-asserted-by":"crossref","first-page":"128","DOI":"10.3389\/fimmu.2017.00128","article-title":"Inhibition of TNF receptor p55 by a domain antibody attenuates the initial phase of acid\u2010induced lung injury in mice","volume":"8","author":"Wilson M. R.","year":"2017","journal-title":"Frontiers in Immunology"},{"key":"e_1_2_7_24_1","doi-asserted-by":"publisher","DOI":"10.1183\/09031936.03.00027502"}],"container-title":["Current Protocols in Mouse Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpmo.85","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cpmo.85","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpmo.85","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,3]],"date-time":"2023-09-03T16:22:02Z","timestamp":1693758122000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpmo.85"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,11,20]]},"references-count":23,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2020,12]]}},"alternative-id":["10.1002\/cpmo.85"],"URL":"https:\/\/doi.org\/10.1002\/cpmo.85","archive":["Portico"],"relation":{},"ISSN":["2161-2617","2161-2617"],"issn-type":[{"value":"2161-2617","type":"print"},{"value":"2161-2617","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,11,20]]},"assertion":[{"value":"2020-11-20","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e85"}}