Current methods for isolating tissue-specific fetal stem cells and deriving primary organoids are limited to samples obtained from pregnancy termination. This study uses single-cell analysis of human amniotic fluid (AF) to identify and isolate viable epithelial stem/progenitor cells of fetal gastrointestinal, renal, and pulmonary origin. These cells formed clonal epithelial organoids exhibiting small intestine, kidney tubule, and lung identities. Lung organoids were also derived from AF and tracheal fluid cells of congenital diaphragmatic hernia (CDH) fetuses, recapitulating some disease features. This approach allows for organoid derivation during ongoing pregnancy, potentially enabling investigation of personalized therapeutic tools and regenerative medicine strategies.

Mattia Francesco Maria Gerli, Giuseppe Calà, Max Arran Beesley, Beatrice Sina, Lucinda Tullie, Kylin Yunyan Sun, Francesco Panariello, Federica Michielin, Joseph R. Davidson, Francesca Maria Russo, Brendan C. Jones, Dani Do Hyang Lee, Savvas Savvidis, Theodoros Xenakis, Ian C. Simcock, Anna A. Straatman-Iwanowska, Robert A. Hirst, Anna L. David, Christopher O’Callaghan, Alessandro Olivo, Simon Eaton, Stavros P. Loukogeorgakis, Davide Cacchiarelli, Jan Deprest, Vivian S. W. Li, Giovanni Giuseppe Giobbe, Paolo De Coppi