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These can be easily translated to human fetal lung samples to allow the investigation of lung development in physiologic and pathologic conditions.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Lungs were harvested from mouse embryos at E12.5 and cultured in three different settings, i.e., whole lung culture, mesenchyme-free\u00a0epithelium culture, and organoid culture. For the whole lung culture, extracted lungs were embedded in Matrigel and incubated on permeable filters. Separately, distal epithelial tips were isolated by firstly\u00a0removing mesothelial and mesenchymal cells, and then severing the tips from the airway tubes. These were then\u00a0cultured either in branch-promoting or self-renewing conditions.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Cultured whole lungs underwent branching morphogenesis similarly to native lungs. Real-time qPCR analysis demonstrated expression of key genes essential for lung bud formation. The culture condition for epithelial tips was optimized by testing different concentrations of FGF10 and CHIR99021 and evaluating branching formation. The epithelial rudiments in self-renewing conditions formed spherical 3D structures with homogeneous Sox9 expression.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>We report efficient protocols for ex vivo culture systems of pseudoglandular stage mouse embryonic lungs. These models can be applied to human samples and could be useful to\u00a0paediatric surgeons to investigate normal lung development, understand the pathogenesis of congenital lung diseases, and explore novel therapeutic strategies.<\/jats:p><\/jats:sec>","DOI":"10.1007\/s00383-021-04864-8","type":"journal-article","created":{"date-parts":[[2021,3,31]],"date-time":"2021-03-31T11:02:51Z","timestamp":1617188571000},"page":"561-568","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["In vitro models of fetal lung development to enhance research into congenital lung diseases"],"prefix":"10.1007","volume":"37","author":[{"given":"Soichi","family":"Shibuya","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica","family":"Allen-Hyttinen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paolo","family":"De Coppi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Federica","family":"Michielin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,3,31]]},"reference":[{"key":"4864_CR1","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1053\/j.sempedsurg.2015.02.003","volume":"24","author":"R Singh","year":"2015","unstructured":"Singh R, Davenport M (2015) The argument for operative approach to asymptomatic lung lesions. 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