{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:14:04Z","timestamp":1785402844135,"version":"3.56.0"},"reference-count":80,"publisher":"Springer Science and Business Media LLC","issue":"3","license":[{"start":{"date-parts":[[2025,8,14]],"date-time":"2025-08-14T00:00:00Z","timestamp":1755129600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.springernature.com\/gp\/researchers\/text-and-data-mining"},{"start":{"date-parts":[[2025,8,14]],"date-time":"2025-08-14T00:00:00Z","timestamp":1755129600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.springernature.com\/gp\/researchers\/text-and-data-mining"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Nat Protoc"],"published-print":{"date-parts":[[2026,3]]},"DOI":"10.1038\/s41596-025-01227-8","type":"journal-article","created":{"date-parts":[[2025,8,14]],"date-time":"2025-08-14T16:03:04Z","timestamp":1755187384000},"page":"1045-1072","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Derivation, expansion and cryopreservation of primary fetal organoids from second and third trimester human amniotic fluid cells"],"prefix":"10.1038","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2297-700X","authenticated-orcid":false,"given":"Giuseppe","family":"Cal\u00e0","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giorgia","family":"D\u2019Ariano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kylin Yunyan","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gloria Ji","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-9360-2626","authenticated-orcid":false,"given":"Giuseppe Matteo","family":"Carrino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandro","family":"Mariani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlotta","family":"Camilli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabella","family":"Fabietti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roberto","family":"Bei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0199-6140","authenticated-orcid":false,"given":"Anna L.","family":"David","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandro Filippo","family":"Pellegata","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2750-8405","authenticated-orcid":false,"given":"Panicos","family":"Shangaris","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3724-6640","authenticated-orcid":false,"given":"Marco","family":"Pellegrini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanni Giuseppe","family":"Giobbe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1659-0207","authenticated-orcid":false,"given":"Paolo","family":"De Coppi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9331-7394","authenticated-orcid":false,"given":"Mattia Francesco Maria","family":"Gerli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,8,14]]},"reference":[{"key":"1227_CR1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s43586-022-00174-y","volume":"2","author":"OZ Zhao","year":"2022","unstructured":"Zhao, O. Z. et al. Organoids. Nat. Rev. Methods Primers 2, 1\u201321 (2022).","journal-title":"Nat. Rev. Methods Primers"},{"key":"1227_CR2","doi-asserted-by":"publisher","first-page":"320","DOI":"10.3389\/fbioe.2023.1058970","volume":"11","author":"G Cal\u00e0","year":"2023","unstructured":"Cal\u00e0, G., Sina, B., De Coppi, P., Giobbe, G. G. & Gerli, M. F. M. Primary human organoids models: current progress and key milestones. Front. Bioeng. Biotechnol. 11, 320 (2023).","journal-title":"Front. Bioeng. Biotechnol."},{"key":"1227_CR3","doi-asserted-by":"publisher","first-page":"1255","DOI":"10.1016\/j.stemcr.2023.05.009","volume":"18","author":"KB Jensen","year":"2023","unstructured":"Jensen, K. B. & Little, M. H. Organoids are not organs: sources of variation and misinformation in organoid biology. Stem Cell Rep. 18, 1255\u20131270 (2023).","journal-title":"Stem Cell Rep."},{"key":"1227_CR4","doi-asserted-by":"publisher","first-page":"3073","DOI":"10.1242\/dev.122820","volume":"142","author":"D Gerrelli","year":"2015","unstructured":"Gerrelli, D., Lisgo, S., Copp, A. J. & Lindsay, S. Enabling research with human embryonic and fetal tissue resources. Development 142, 3073\u20133076 (2015).","journal-title":"Development"},{"key":"1227_CR5","unstructured":"Abortion care. RCOG https:\/\/www.rcog.org.uk\/for-the-public\/browse-our-patient-information\/abortion-care\/ (2012)."},{"key":"1227_CR6","unstructured":"Fetal tissue research: a weapon and a casualty in the war against abortion. Guttmacher Institute https:\/\/www.guttmacher.org\/gpr\/2016\/fetal-tissue-research-weapon-and-casualty-war-against-abortion (2016)."},{"key":"1227_CR7","doi-asserted-by":"publisher","first-page":"777","DOI":"10.1016\/j.stemcr.2019.10.003","volume":"13","author":"JM McCune","year":"2019","unstructured":"McCune, J. M. & Weissman, I. L. The ban on US government funding research using human fetal tissues: how does this fit with the nih mission to advance medical science for the benefit of the citizenry? Stem Cell Rep. 13, 777\u2013786 (2019).","journal-title":"Stem Cell Rep."},{"key":"1227_CR8","doi-asserted-by":"publisher","first-page":"2300","DOI":"10.1016\/j.stemcr.2023.10.008","volume":"18","author":"J Brumbaugh","year":"2023","unstructured":"Brumbaugh, J., Aguado, B. A., Lysaght, T. & Goldstein, L. S. B. Human fetal tissue is critical for biomedical research. Stem Cell Rep. 18, 2300\u20132312 (2023).","journal-title":"Stem Cell Rep."},{"key":"1227_CR9","doi-asserted-by":"publisher","first-page":"875","DOI":"10.1038\/s41591-024-02807-z","volume":"30","author":"MFM Gerli","year":"2024","unstructured":"Gerli, M. F. M. et al. Single-cell guided prenatal derivation of primary fetal epithelial organoids from human amniotic and tracheal fluids. Nat. Med. 30, 875\u2013887 (2024).","journal-title":"Nat. Med."},{"key":"1227_CR10","doi-asserted-by":"publisher","first-page":"341","DOI":"10.1038\/sj.jp.7211290","volume":"25","author":"MA Underwood","year":"2005","unstructured":"Underwood, M. A., Gilbert, W. M. & Sherman, M. P. Amniotic fluid: not just fetal urine anymore. J. Perinat. 25, 341\u2013348 (2005).","journal-title":"J. Perinat."},{"key":"1227_CR11","doi-asserted-by":"publisher","unstructured":"Beall, M. H., van den Wijngaard, J. P. H. M., van Gemert, M. & Ross, M. G. in Nephrology and Fluid\/Electrolyte Physiology 3\u201318 (Springer, 2019); https:\/\/doi.org\/10.1016\/B978-0-323-53367-6.00001-7.","DOI":"10.1016\/B978-0-323-53367-6.00001-7"},{"key":"1227_CR12","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2027030","author":"JA Deprest","year":"2021","unstructured":"Deprest, J. A. et al. Randomized trial of fetal surgery for severe left diaphragmatic hernia. N. Engl. J. Med. https:\/\/doi.org\/10.1056\/NEJMoa2027030 (2021).","journal-title":"N. Engl. J. Med."},{"key":"1227_CR13","doi-asserted-by":"crossref","unstructured":"Deprest, J. A. et al. Randomized trial of fetal surgery for moderate left diaphragmatic hernia. New Engl. J. Med. 385, 119\u2013129 (2021).","DOI":"10.1056\/NEJMoa2026983"},{"key":"1227_CR14","doi-asserted-by":"publisher","first-page":"100","DOI":"10.1038\/nbt1274","volume":"25","author":"P De Coppi","year":"2007","unstructured":"De Coppi, P. et al. Isolation of amniotic stem cell lines with potential for therapy. Nat. Biotechnol. 25, 100\u2013106 (2007).","journal-title":"Nat. Biotechnol."},{"key":"1227_CR15","doi-asserted-by":"publisher","first-page":"136","DOI":"10.1056\/NEJMoa032597","volume":"351","author":"M-V Senat","year":"2004","unstructured":"Senat, M.-V. et al. Endoscopic laser surgery versus serial amnioreduction for severe twin-to-twin transfusion syndrome. N. Engl. J. Med. 351, 136\u2013144 (2004).","journal-title":"N. Engl. J. Med."},{"key":"1227_CR16","doi-asserted-by":"publisher","first-page":"993","DOI":"10.1056\/NEJMoa1014379","volume":"364","author":"NS Adzick","year":"2011","unstructured":"Adzick, N. S. et al. A randomized trial of prenatal versus postnatal repair of myelomeningocele. N. Engl. J. Med. 364, 993\u20131004 (2011).","journal-title":"N. Engl. J. Med."},{"key":"1227_CR17","doi-asserted-by":"publisher","first-page":"4841","DOI":"10.1016\/j.cell.2022.11.005","volume":"185","author":"P He","year":"2022","unstructured":"He, P. et al. A human fetal lung cell atlas uncovers proximal-distal gradients of differentiation and key regulators of epithelial fates. Cell 185, 4841\u20134860.e25 (2022).","journal-title":"Cell"},{"key":"1227_CR18","doi-asserted-by":"crossref","unstructured":"Lim, K. et al. Organoid modeling of human fetal lung alveolar development reveals mechanisms of cell fate patterning and neonatal respiratory disease. Cell Stem Cell 30, 20\u201337.e9 (2022).","DOI":"10.1016\/j.stem.2022.11.013"},{"key":"1227_CR19","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1136\/gutjnl-2017-314817","volume":"68","author":"J Kraiczy","year":"2019","unstructured":"Kraiczy, J. et al. DNA methylation defines regional identity of human intestinal epithelial organoids and undergoes dynamic changes during development. Gut 68, 49\u201361 (2019).","journal-title":"Gut"},{"key":"1227_CR20","doi-asserted-by":"publisher","first-page":"771","DOI":"10.1016\/j.devcel.2020.11.010","volume":"55","author":"R Elmentaite","year":"2020","unstructured":"Elmentaite, R. et al. Single-cell sequencing of developing human gut reveals transcriptional links to childhood Crohn\u2019s disease. Dev. Cell 55, 771\u2013783.e5 (2020).","journal-title":"Dev. Cell"},{"key":"1227_CR21","doi-asserted-by":"publisher","first-page":"1487","DOI":"10.1038\/s41556-022-00989-7","volume":"24","author":"BT Wesley","year":"2022","unstructured":"Wesley, B. T. et al. Single-cell atlas of human liver development reveals pathways directing hepatic cell fates. Nat. Cell Biol. 24, 1487\u20131498 (2022).","journal-title":"Nat. Cell Biol."},{"key":"1227_CR22","doi-asserted-by":"publisher","unstructured":"Andersson-Rolf, A. et al. Long-term in vitro expansion of a human fetal pancreas stem cell that generates all three pancreatic cell lineages. Cell https:\/\/doi.org\/10.1016\/j.cell.2024.10.044 (2024).","DOI":"10.1016\/j.cell.2024.10.044"},{"key":"1227_CR23","doi-asserted-by":"publisher","first-page":"117","DOI":"10.1016\/j.devcel.2020.01.033","volume":"53","author":"AJ Miller","year":"2020","unstructured":"Miller, A. J. et al. In vitro and in vivo development of the human airway at single-cell resolution. Dev. Cell 53, 117\u2013128.e6 (2020).","journal-title":"Dev. Cell"},{"key":"1227_CR24","doi-asserted-by":"publisher","first-page":"734","DOI":"10.1016\/j.stem.2013.09.015","volume":"13","author":"RP Fordham","year":"2013","unstructured":"Fordham, R. P. et al. Transplantation of expanded fetal intestinal progenitors contributes to colon regeneration after injury. Cell Stem Cell 13, 734\u2013744 (2013).","journal-title":"Cell Stem Cell"},{"key":"1227_CR25","doi-asserted-by":"publisher","first-page":"3441","DOI":"10.1038\/s41596-020-0381-x","volume":"15","author":"MA Sheridan","year":"2020","unstructured":"Sheridan, M. A. et al. Establishment and differentiation of long-term trophoblast organoid cultures from the human placenta. Nat. Protoc. 15, 3441\u20133463 (2020).","journal-title":"Nat. Protoc."},{"key":"1227_CR26","doi-asserted-by":"publisher","DOI":"10.1002\/ADVS.202105568","author":"J Liang","year":"2022","unstructured":"Liang, J. et al. Modeling human thyroid development by fetal tissue-derived organoid culture. Adv. Sci. https:\/\/doi.org\/10.1002\/ADVS.202105568 (2022).","journal-title":"Adv. Sci."},{"key":"1227_CR27","doi-asserted-by":"publisher","unstructured":"Minimally invasive derivation of primary human epithelial organoids from fetal fluids. Nat. Med. https:\/\/doi.org\/10.1038\/s41591-024-02831-z (2024).","DOI":"10.1038\/s41591-024-02831-z"},{"key":"1227_CR28","doi-asserted-by":"publisher","first-page":"32","DOI":"10.1038\/s44222-022-00013-5","volume":"1","author":"MH Geurts","year":"2023","unstructured":"Geurts, M. H. & Clevers, H. CRISPR engineering in organoids for gene repair and disease modelling. Nat. Rev. Bioeng. 1, 32\u201345 (2023).","journal-title":"Nat. Rev. Bioeng."},{"key":"1227_CR29","doi-asserted-by":"publisher","first-page":"653","DOI":"10.1016\/j.stem.2013.11.002","volume":"13","author":"G Schwank","year":"2013","unstructured":"Schwank, G. et al. Functional repair of CFTR by CRISPR\/Cas9 in intestinal stem cell organoids of cystic fibrosis patients. Cell Stem Cell 13, 653\u2013658 (2013).","journal-title":"Cell Stem Cell"},{"key":"1227_CR30","doi-asserted-by":"publisher","first-page":"431","DOI":"10.1016\/j.stem.2020.02.007","volume":"26","author":"T Ringel","year":"2020","unstructured":"Ringel, T. et al. Genome-scale CRISPR screening in human intestinal organoids identifies drivers of TGF-\u03b2 resistance. Cell Stem Cell 26, 431\u2013440.e8 (2020).","journal-title":"Cell Stem Cell"},{"key":"1227_CR31","doi-asserted-by":"publisher","unstructured":"Pagliaro, A., Andreatta, F., Finger, R., Artegiani, B. & Hendriks, D. Generation of human fetal brain organoids and their CRISPR engineering for brain tumor modeling. Nat. Protoc. https:\/\/doi.org\/10.1038\/s41596-024-01107-7 (2025).","DOI":"10.1038\/s41596-024-01107-7"},{"key":"1227_CR32","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1016\/j.devcel.2025.01.005","volume":"60","author":"B Artegiani","year":"2025","unstructured":"Artegiani, B. & Hendriks, D. Organoids from pluripotent stem cells and human tissues: when two cultures meet each other. Dev. Cell 60, 493\u2013511 (2025).","journal-title":"Dev. Cell"},{"key":"1227_CR33","doi-asserted-by":"publisher","first-page":"639","DOI":"10.1038\/s44318-024-00328-6","volume":"44","author":"K Lim","year":"2025","unstructured":"Lim, K. et al. A novel human fetal lung-derived alveolar organoid model reveals mechanisms of surfactant protein C maturation relevant to interstitial lung disease. EMBO J. 44, 639\u2013664 (2025).","journal-title":"EMBO J."},{"key":"1227_CR34","doi-asserted-by":"publisher","first-page":"3380","DOI":"10.1038\/s41596-020-0379-4","volume":"15","author":"E Driehuis","year":"2020","unstructured":"Driehuis, E., Kretzschmar, K. & Clevers, H. Establishment of patient-derived cancer organoids for drug-screening applications. Nat. Protoc. 15, 3380\u20133409 (2020).","journal-title":"Nat. Protoc."},{"key":"1227_CR35","doi-asserted-by":"publisher","first-page":"1041","DOI":"10.1038\/s41556-019-0360-z","volume":"21","author":"M Boretto","year":"2019","unstructured":"Boretto, M. et al. Patient-derived organoids from endometrial disease capture clinical heterogeneity and are amenable to drug screening. Nat. Cell Biol. 21, 1041\u20131051 (2019).","journal-title":"Nat. Cell Biol."},{"key":"1227_CR36","doi-asserted-by":"publisher","first-page":"109218","DOI":"10.1016\/j.celrep.2021.109218","volume":"35","author":"T Ebisudani","year":"2021","unstructured":"Ebisudani, T. et al. Direct derivation of human alveolospheres for SARS-CoV-2 infection modeling and drug screening. Cell Rep. 35, 109218 (2021).","journal-title":"Cell Rep."},{"key":"1227_CR37","doi-asserted-by":"publisher","first-page":"571","DOI":"10.1038\/s41580-020-0259-3","volume":"21","author":"J Kim","year":"2020","unstructured":"Kim, J., Koo, B. K. & Knoblich, J. A. Human organoids: model systems for human biology and medicine. Nat. Rev. Mol. Cell Biol. 21, 571\u2013584 (2020).","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"1227_CR38","doi-asserted-by":"publisher","first-page":"108415","DOI":"10.1016\/j.envint.2024.108415","volume":"184","author":"C Hu","year":"2024","unstructured":"Hu, C. et al. Organoids and organoids-on-a-chip as the new testing strategies for environmental toxicology\u2014applications & advantages. Environ. Int. 184, 108415 (2024).","journal-title":"Environ. Int."},{"key":"1227_CR39","doi-asserted-by":"publisher","first-page":"851","DOI":"10.1039\/C7LC01084B","volume":"18","author":"Y Wang","year":"2018","unstructured":"Wang, Y., Wang, L., Zhu, Y. & Qin, J. Human brain organoid-on-a-chip to model prenatal nicotine exposure. Lab Chip 18, 851\u2013860 (2018).","journal-title":"Lab Chip"},{"key":"1227_CR40","doi-asserted-by":"publisher","first-page":"107200","DOI":"10.1016\/j.envint.2022.107200","volume":"163","author":"AS Winkler","year":"2022","unstructured":"Winkler, A. S. et al. Human airway organoids and microplastic fibers: a new exposure model for emerging contaminants. Environ. Int. 163, 107200 (2022).","journal-title":"Environ. Int."},{"key":"1227_CR41","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1038\/s41591-024-03489-3","volume":"31","author":"MMA Verstegen","year":"2025","unstructured":"Verstegen, M. M. A. et al. Clinical applications of human organoids. Nat. Med. 31, 409\u2013421 (2025).","journal-title":"Nat. Med."},{"key":"1227_CR42","doi-asserted-by":"publisher","first-page":"e100300","DOI":"10.15252\/embj.2018100300","volume":"38","author":"N Sachs","year":"2019","unstructured":"Sachs, N. et al. Long-term expanding human airway organoids for disease modeling. EMBO J. 38, e100300 (2019).","journal-title":"EMBO J."},{"key":"1227_CR43","doi-asserted-by":"publisher","first-page":"303","DOI":"10.1038\/s41587-019-0048-8","volume":"37","author":"F Schutgens","year":"2019","unstructured":"Schutgens, F. et al. Tubuloids derived from human adult kidney and urine for personalized disease modeling. Nat. Biotechnol. 37, 303\u2013313 (2019).","journal-title":"Nat. Biotechnol."},{"key":"1227_CR44","doi-asserted-by":"publisher","unstructured":"Roos, F. J. M. et al. Human bile contains cholangiocyte organoid-initiating cells which expand as functional cholangiocytes in non-canonical Wnt stimulating conditions. Front. Cell Dev. Biol. https:\/\/doi.org\/10.3389\/fcell.2020.630492 (2021).","DOI":"10.3389\/fcell.2020.630492"},{"key":"1227_CR45","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s42003-021-02194-y","volume":"4","author":"T Cindrova-Davies","year":"2021","unstructured":"Cindrova-Davies, T. et al. Menstrual flow as a non-invasive source of endometrial organoids. Commun. Biol. 4, 1\u20138 (2021).","journal-title":"Commun. Biol."},{"key":"1227_CR46","first-page":"1","volume":"14","author":"X-Z Huang","year":"2023","unstructured":"Huang, X.-Z. et al. Single-cell sequencing of ascites fluid illustrates heterogeneity and therapy-induced evolution during gastric cancer peritoneal metastasis. Nat. Commun. 14, 1\u201322 (2023).","journal-title":"Nat. Commun."},{"key":"1227_CR47","doi-asserted-by":"publisher","first-page":"963689722112448","DOI":"10.1177\/09636897221124481","volume":"31","author":"J Liang","year":"2022","unstructured":"Liang, J., Li, X., Dong, Y. & Zhao, B. Modeling human organ development and diseases with fetal tissue-derived organoids. Cell Transplant. 31, 9636897221124481 (2022).","journal-title":"Cell Transplant."},{"key":"1227_CR48","doi-asserted-by":"publisher","first-page":"196","DOI":"10.1038\/s41586-021-03620-1","volume":"597","author":"M Haniffa","year":"2021","unstructured":"Haniffa, M. et al. A roadmap for the Human Developmental Cell Atlas. Nature 597, 196\u2013205 (2021).","journal-title":"Nature"},{"key":"1227_CR49","doi-asserted-by":"publisher","first-page":"643","DOI":"10.1016\/S2352-4642(22)00192-4","volume":"6","author":"P de Coppi","year":"2022","unstructured":"de Coppi, P. et al. Regenerative medicine: prenatal approaches. Lancet Child Adolesc. Health 6, 643\u2013653 (2022).","journal-title":"Lancet Child Adolesc. Health"},{"key":"1227_CR50","doi-asserted-by":"publisher","first-page":"273","DOI":"10.1016\/j.ydbio.2017.05.015","volume":"428","author":"K Kretzschmar","year":"2017","unstructured":"Kretzschmar, K. & Clevers, H. Wnt\/\u03b2-catenin signaling in adult mammalian epithelial stem cells. Dev. Biol. 428, 273\u2013282 (2017).","journal-title":"Dev. Biol."},{"key":"1227_CR51","doi-asserted-by":"publisher","first-page":"1586","DOI":"10.1016\/j.cell.2016.05.082","volume":"165","author":"H Clevers","year":"2016","unstructured":"Clevers, H. Modeling development and disease with organoids. Cell 165, 1586\u20131597 (2016).","journal-title":"Cell"},{"key":"1227_CR52","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1016\/j.stem.2023.12.006","volume":"31","author":"J Beumer","year":"2024","unstructured":"Beumer, J. & Clevers, H. Hallmarks of stemness in mammalian tissues. Cell Stem Cell 31, 7\u201324 (2024).","journal-title":"Cell Stem Cell"},{"key":"1227_CR53","doi-asserted-by":"publisher","first-page":"671","DOI":"10.1038\/s41576-018-0051-9","volume":"19","author":"G Rossi","year":"2018","unstructured":"Rossi, G., Manfrin, A. & Lutolf, M. P. Progress and potential in organoid research. Nat. Rev. Genet. 19, 671\u2013687 (2018).","journal-title":"Nat. Rev. Genet."},{"key":"1227_CR54","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s13073-019-0615-0","volume":"11","author":"AN Combes","year":"2019","unstructured":"Combes, A. N., Zappia, L., Er, P. X., Oshlack, A. & Little, M. H. Single-cell analysis reveals congruence between kidney organoids and human fetal kidney. Genome Med. 11, 1\u201315 (2019).","journal-title":"Genome Med."},{"key":"1227_CR55","doi-asserted-by":"publisher","first-page":"e1","DOI":"10.1111\/1471-0528.16821","volume":"129","author":"K Navaratnam","year":"2022","unstructured":"Navaratnam, K. & Alfirevic, Z. Amniocentesis and chorionic villus sampling. BJOG 129, e1\u2013e15 (2022).","journal-title":"BJOG"},{"key":"1227_CR56","doi-asserted-by":"publisher","unstructured":"Cao, J. et al. A human cell atlas of fetal gene expression. Science https:\/\/doi.org\/10.1126\/science.aba772 (2020).","DOI":"10.1126\/science.aba772"},{"key":"1227_CR57","doi-asserted-by":"publisher","first-page":"661","DOI":"10.1038\/s41592-018-0113-0","volume":"15","author":"JG Camp","year":"2018","unstructured":"Camp, J. G., Wollny, D. & Treutlein, B. Single-cell genomics to guide human stem cell and tissue engineering. Nat. Methods 15, 661\u2013667 (2018).","journal-title":"Nat. Methods"},{"key":"1227_CR58","doi-asserted-by":"publisher","first-page":"2175","DOI":"10.1056\/NEJMoa1203382","volume":"367","author":"RJ Wapner","year":"2012","unstructured":"Wapner, R. J. et al. Chromosomal microarray versus karyotyping for prenatal diagnosis. N. Engl. J. Med. 367, 2175\u20132184 (2012).","journal-title":"N. Engl. J. Med."},{"key":"1227_CR59","doi-asserted-by":"publisher","first-page":"102575","DOI":"10.1016\/j.bpobgyn.2024.102575","volume":"98","author":"MA Shear","year":"2025","unstructured":"Shear, M. A., Robinson, P. N. & Sparks, T. N. Fetal imaging, phenotyping, and genomic testing in modern prenatal diagnosis. Best. Pract. Res. Clin. Obstet. Gynaecol. 98, 102575 (2025).","journal-title":"Best. Pract. Res. Clin. Obstet. Gynaecol."},{"key":"1227_CR60","doi-asserted-by":"publisher","first-page":"816","DOI":"10.1016\/j.placenta.2006.11.009","volume":"28","author":"MH Beall","year":"2007","unstructured":"Beall, M. H., van den Wijngaard, J. P. H. M., van Gemert, M. J. C. & Ross, M. G. Amniotic fluid water dynamics. Placenta 28, 816\u2013823 (2007).","journal-title":"Placenta"},{"key":"1227_CR61","doi-asserted-by":"publisher","first-page":"761","DOI":"10.1387\/ijdb.092935md","volume":"54","author":"MP Dobreva","year":"2010","unstructured":"Dobreva, M. P., Pereira, P. N. G., Deprest, J. & Zwijsen, A. On the origin of amniotic stem cells: of mice and men. Int. J. Dev. Biol. 54, 761\u2013777 (2010).","journal-title":"Int. J. Dev. Biol."},{"key":"1227_CR62","doi-asserted-by":"publisher","first-page":"348","DOI":"10.1093\/oxfordjournals.bmb.a071847","volume":"39","author":"CM Gosden","year":"1983","unstructured":"Gosden, C. M. Amniotic fluid cell types and culture. Br. Med. Bull. 39, 348\u2013354 (1983).","journal-title":"Br. Med. Bull."},{"key":"1227_CR63","doi-asserted-by":"publisher","first-page":"262","DOI":"10.1038\/nature07935","volume":"459","author":"T Sato","year":"2009","unstructured":"Sato, T. et al. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459, 262\u2013265 (2009).","journal-title":"Nature"},{"key":"1227_CR64","doi-asserted-by":"publisher","first-page":"e106","DOI":"10.1002\/cpim.106","volume":"130","author":"C Pleguezuelos-Manzano","year":"2020","unstructured":"Pleguezuelos-Manzano, C. et al. Establishment and culture of human intestinal organoids derived from adult stem cells. Curr. Protoc. Immunol. 130, e106 (2020).","journal-title":"Curr. Protoc. Immunol."},{"key":"1227_CR65","doi-asserted-by":"publisher","first-page":"260","DOI":"10.1038\/nature19768","volume":"538","author":"F Blokzijl","year":"2016","unstructured":"Blokzijl, F. et al. Tissue-specific mutation accumulation in human adult stem cells during life. Nature 538, 260\u2013264 (2016).","journal-title":"Nature"},{"key":"1227_CR66","doi-asserted-by":"publisher","first-page":"457","DOI":"10.1038\/s41586-018-0024-3","volume":"556","author":"SF Roerink","year":"2018","unstructured":"Roerink, S. F. et al. Intra-tumour diversification in colorectal cancer at the single-cell level. Nature 556, 457\u2013462 (2018).","journal-title":"Nature"},{"key":"1227_CR67","doi-asserted-by":"publisher","unstructured":"Meran, L., Tullie, L., Eaton, S., De Coppi, P. & Li, V. S. W. Bioengineering human intestinal mucosal grafts using patient-derived organoids, fibroblasts and scaffolds. Nat. Protoc. https:\/\/doi.org\/10.1038\/s41596-022-00751-1 (2022).","DOI":"10.1038\/s41596-022-00751-1"},{"key":"1227_CR68","doi-asserted-by":"publisher","first-page":"588","DOI":"10.1016\/j.devcel.2009.02.010","volume":"16","author":"N Gao","year":"2009","unstructured":"Gao, N., White, P. & Kaestner, K. H. Establishment of intestinal identity and epithelial\u2013mesenchymal signaling by Cdx2. Dev. Cell 16, 588 (2009).","journal-title":"Dev. Cell"},{"key":"1227_CR69","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1016\/j.ydbio.2009.12.025","volume":"339","author":"S Grainger","year":"2010","unstructured":"Grainger, S., Savory, J. G. A. & Lohnes, D. Cdx2 regulates patterning of the intestinal epithelium. Dev. Biol. 339, 155\u2013165 (2010).","journal-title":"Dev. Biol."},{"key":"1227_CR70","doi-asserted-by":"publisher","first-page":"dev172189","DOI":"10.1242\/dev.172189","volume":"146","author":"N Kumar","year":"2019","unstructured":"Kumar, N. et al. The lineage-specific transcription factor CDX2 navigates dynamic chromatin to control distinct stages of intestine development. Development 146, dev172189 (2019).","journal-title":"Development"},{"key":"1227_CR71","doi-asserted-by":"publisher","first-page":"2958","DOI":"10.1101\/gad.240102","volume":"16","author":"M Bouchard","year":"2002","unstructured":"Bouchard, M., Souabni, A., Mandler, M., Neub\u00fcser, A. & Busslinger, M. Nephric lineage specification by Pax2 and Pax8. Genes Dev. 16, 2958\u20132970 (2002).","journal-title":"Genes Dev."},{"key":"1227_CR72","doi-asserted-by":"publisher","first-page":"1121","DOI":"10.1681\/ASN.2006070739","volume":"18","author":"M Narlis","year":"2007","unstructured":"Narlis, M., Grote, D., Gaitan, Y., Boualia, S. K. & Bouchard, M. Pax2 and Pax8 regulate branching morphogenesis and nephron differentiation in the developing kidney. J. Am. Soc. Nephrol. 18, 1121\u20131129 (2007).","journal-title":"J. Am. Soc. Nephrol."},{"key":"1227_CR73","doi-asserted-by":"publisher","first-page":"1093","DOI":"10.1242\/dev.113.4.1093","volume":"113","author":"D Lazzaro","year":"1991","unstructured":"Lazzaro, D., Price, M., De Felice, M. & Di Lauro, R. The transcription factor TTF-1 is expressed at the onset of thyroid and lung morphogenesis and in restricted regions of the foetal brain. Development 113, 1093\u20131104 (1991).","journal-title":"Development"},{"key":"1227_CR74","doi-asserted-by":"publisher","first-page":"694","DOI":"10.1006\/dbio.1995.8080","volume":"172","author":"P Minoo","year":"1995","unstructured":"Minoo, P. et al. TTF-1 regulates lung epithelial morphogenesis. Dev. Biol. 172, 694\u2013698 (1995).","journal-title":"Dev. Biol."},{"key":"1227_CR75","doi-asserted-by":"publisher","first-page":"2277","DOI":"10.1172\/JCI89950","volume":"127","author":"F Hawkins","year":"2017","unstructured":"Hawkins, F. et al. Prospective isolation of NKX2-1\u2013expressing human lung progenitors derived from pluripotent stem cells. J. Clin. Invest. 127, 2277\u20132294 (2017).","journal-title":"J. Clin. Invest."},{"key":"1227_CR76","doi-asserted-by":"publisher","first-page":"1936","DOI":"10.1038\/s41596-020-00474-1","volume":"16","author":"JF Dekkers","year":"2021","unstructured":"Dekkers, J. F. et al. Long-term culture, genetic manipulation and xenotransplantation of human normal and breast cancer organoids. Nat. Protoc. 16, 1936\u20131965 (2021).","journal-title":"Nat. Protoc."},{"key":"1227_CR77","doi-asserted-by":"publisher","first-page":"e2300543","DOI":"10.1002\/biot.202300543","volume":"19","author":"H Han","year":"2024","unstructured":"Han, H., Zhan, T., Guo, N., Cui, M. & Xu, Y. Cryopreservation of organoids: Strategies, innovation, and future prospects. Biotechnol. J. 19, e2300543 (2024).","journal-title":"Biotechnol. J."},{"key":"1227_CR78","doi-asserted-by":"publisher","first-page":"100777","DOI":"10.1016\/j.crmeth.2024.100777","volume":"4","author":"W Xue","year":"2024","unstructured":"Xue, W. et al. Effective cryopreservation of human brain tissue and neural organoids. Cell Rep. Methods 4, 100777 (2024).","journal-title":"Cell Rep. Methods"},{"key":"1227_CR79","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/s00018-024-05352-7","volume":"81","author":"P Mashouf","year":"2024","unstructured":"Mashouf, P., Tabibzadeh, N., Kuraoka, S., Oishi, H. & Morizane, R. Cryopreservation of human kidney organoids. Cell. Mol. Life Sci. 81, 1\u201323 (2024).","journal-title":"Cell. Mol. Life Sci."},{"key":"1227_CR80","doi-asserted-by":"publisher","first-page":"1756","DOI":"10.1038\/s41596-019-0160-8","volume":"14","author":"JF Dekkers","year":"2019","unstructured":"Dekkers, J. F. et al. High-resolution 3D imaging of fixed and cleared organoids. Nat. Protoc. 14, 1756\u20131771 (2019).","journal-title":"Nat. Protoc."}],"container-title":["Nature Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41596-025-01227-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41596-025-01227-8","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41596-025-01227-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,11]],"date-time":"2026-03-11T15:01:37Z","timestamp":1773241297000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41596-025-01227-8"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,14]]},"references-count":80,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2026,3]]}},"alternative-id":["1227"],"URL":"https:\/\/doi.org\/10.1038\/s41596-025-01227-8","relation":{},"ISSN":["1754-2189","1750-2799"],"issn-type":[{"value":"1754-2189","type":"print"},{"value":"1750-2799","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,8,14]]},"assertion":[{"value":"31 January 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"6 June 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"14 August 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"M.F.M.G., G.G.G., P.D.C. and G.C. are inventors of the patent \u2018Derivation of Primary Organoids from the Fetal Fluids\u2019 filed by UCL on 18 April 2023, with application number GB2305703.7.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}]}}