The study addresses the limitation in obtaining tissue-specific fetal stem cells for prenatal investigations due to the current reliance on samples from terminated pregnancies. To overcome this hurdle, the researchers aimed to isolate and expand fetal stem cells directly during pregnancy, without the need for tissue samples or reprogramming. Amniotic fluid (AF) was identified as a source of cells representing various developing organs. Through single-cell analysis, viable epithelial stem/progenitor cells from fetal gastrointestinal, renal, and pulmonary origins were isolated. These cells, when cultured, formed clonal epithelial organoids representing the small intestine, kidney tubule, and lung. AF organoids exhibited characteristics, including transcriptomic, protein expression, and functional features, specific to their respective tissues of origin. The researchers also derived lung organoids from AF and tracheal fluid cells of fetuses with congenital diaphragmatic hernia, successfully replicating some disease features. The timeline for AF organoid derivation is compatible with prenatal intervention, offering a potential avenue for investigating therapeutic tools and personalized regenerative medicine strategies for fetuses at clinically relevant developmental stages.