02864nas a2200505   4500000000100000000000100001008004100002260001500043653003500058653002600093653001500119653002300134100003300157700001900190700002200209700001800231700001900249700002100268700002500289700002300314700002300337700002600360700002100386700002200407700002000429700002200449700001900471700003200490700002000522700001800542700003000560700002100590700001600611700003000627700002400657700001600681700002000697700002900717700001900746245012100765856005500886300000900941520139400950022001402344       2024                            d  c2024-03-0410aExperimental models of disease10aRegenerative Medicine10aStem cells10aStem-cell research1 aMattia Francesco Maria Gerli1 aGiuseppe Calà1 aMax Arran Beesley1 aBeatrice Sina1 aLucinda Tullie1 aKylin Yunyan Sun1 aFrancesco Panariello1 aFederica Michielin1 aJoseph R. Davidson1 aFrancesca Maria Russo1 aBrendan C. Jones1 aDani Do Hyang Lee1 aSavvas Savvidis1 aTheodoros Xenakis1 aIan C. Simcock1 aAnna A. Straatman-Iwanowska1 aRobert A. Hirst1 aAnna L. David1 aChristopher O’Callaghan1 aAlessandro Olivo1 aSimon Eaton1 aStavros P. Loukogeorgakis1 aDavide Cacchiarelli1 aJan Deprest1 aVivian S. W. Li1 aGiovanni Giuseppe Giobbe1 aPaolo De Coppi00aSingle-cell guided prenatal derivation of primary fetal epithelial organoids from human amniotic and tracheal fluids  uhttps://www.nature.com/articles/s41591-024-02807-z  a1-133 aIsolation of tissue-specific fetal stem cells and derivation of primary organoids is limited to samples obtained from termination of pregnancies, hampering prenatal investigation of fetal development and congenital diseases. Therefore, new patient-specific in vitro models are needed. To this aim, isolation and expansion of fetal stem cells during pregnancy, without the need for tissue samples or reprogramming, would be advantageous. Amniotic fluid (AF) is a source of cells from multiple developing organs. Using single-cell analysis, we characterized the cellular identities present in human AF. We identified and isolated viable epithelial stem/progenitor cells of fetal gastrointestinal, renal and pulmonary origin. Upon culture, these cells formed clonal epithelial organoids, manifesting small intestine, kidney tubule and lung identity. AF organoids exhibit transcriptomic, protein expression and functional features of their tissue of origin. With relevance for prenatal disease modeling, we derived lung organoids from AF and tracheal fluid cells of congenital diaphragmatic hernia fetuses, recapitulating some features of the disease. AF organoids are derived in a timeline compatible with prenatal intervention, potentially allowing investigation of therapeutic tools and regenerative medicine strategies personalized to the fetus at clinically relevant developmental stages.  a1546-170X