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Here, we present a simple and reliable minimally invasive transthoracic intrapleural injection technique that could improve the efficiency of orthotopic PM model generation. By incorporating a simple needle sleeve to control the injection depth, this method eliminates the need for surgery or general anaesthesia, reducing technical complexity and animal stress while ensuring precise delivery into the pleural cavity. We demonstrate the effectiveness of this approach by achieving a 100% tumour engraftment rate following the injection of AE17 tumour cells. Additionally, this technique has been successfully used for asbestos fibre injection in mesothelioma models, highlighting its versatility. By providing a more accessible, standardised alternative to existing methods, this protocol improves the reliability of PM models and facilitates broader adoption by researchers, including those with limited experience in invasive procedures.<\/jats:p>","DOI":"10.3390\/mps8030055","type":"journal-article","created":{"date-parts":[[2025,5,28]],"date-time":"2025-05-28T03:52:08Z","timestamp":1748404328000},"page":"55","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["A Minimally Invasive Transthoracic Injection Technique for Reproducible Intrapleural Delivery in Mice"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4936-859X","authenticated-orcid":false,"given":"Sophie","family":"Rovers","sequence":"first","affiliation":[{"name":"Center for Oncological Research (CORE), University of Antwerp, 2000 Antwerpen, 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(CORE), University of Antwerp, 2000 Antwerpen, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dieter","family":"Peeters","sequence":"additional","affiliation":[{"name":"Department of Pathology, Antwerp University Hospital, 2650 Edegem, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Danielle","family":"McKinven","sequence":"additional","affiliation":[{"name":"School of Cancer Sciences, University of Glasgow, Glasgow G61 1BD, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Doig","sequence":"additional","affiliation":[{"name":"School of Cancer Sciences, University of Glasgow, Glasgow G61 1BD, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7174-4144","authenticated-orcid":false,"given":"Filip","family":"Lardon","sequence":"additional","affiliation":[{"name":"Center for Oncological Research (CORE), University of Antwerp, 2000 Antwerpen, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7872-8622","authenticated-orcid":false,"given":"Jan","family":"van Meerbeeck","sequence":"additional","affiliation":[{"name":"Multidisciplinary Oncological Center Antwerp (MOCA), Antwerp University Hospital, 2650 Edegem, Belgium"},{"name":"European Reference Network for Rare or Low Prevalence Lung Diseases\u2014ERN-LUNG, 60596 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3173-0500","authenticated-orcid":false,"given":"Elly","family":"Marcq","sequence":"additional","affiliation":[{"name":"Laboratory of Dendritic Cell Biology and Cancer Immunotherapy, VIB Center for Inflammation Research, 9052 Gent, Belgium"},{"name":"Brussels Center for Immunology, Vrije Universiteit Brussel, 1050 Ixelles, 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