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Figure 3. Characterization of primary fetal epithelial amniotic fluid organoids (AFOs). Phase-contrast images show organoid formation from amniotic
fluid cells, with varying morphologies observed by day 14. Organoid formation efficiency and size at P0 are quantified, with minimal correlation to
gestational age. X-ray PC-CT reveals compact and cystic phenotypes. Immunofluorescence at P3 confirms proliferation (Ki67), epithelial marker
expression (EpCAM, ECAD, pan-cytokeratin), and polarization (ZO-1, ITGβ4). PCA and scRNA-seq identify distinct clusters corresponding to
organoids from various fetal tissues. Copyright © 2024 The author(s).
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Abbreviations: GA: Gestational age; PC-CT: Photon-counting computed tomography; PCA: Principal component analysis; scRNA-seq: Single-cell RNA
sequencing.
characteristics of intestinal epithelial cells and digestive of fetuses with congenital diaphragmatic hernia (CDH).
enzyme activity upon maturation. Kidney organoids These organoids exhibited disease-related features of
(kidney AF organoids, KAFOs) showed features of CDH, such as increased expression of SOX9 and changes
renal tubule cells and displayed functional voltage-gated in the expression of surfactant-related genes. The study
potassium channels and tight junctions. Lung organoids provides a novel approach to obtaining fetal stem cells
(lung AF organoids, LAFOs) demonstrated traits of from AF and tracheal fluid and to culture tissue-specific
lung epithelial cells, such as normal cilia structure and organoids without the need to terminate the pregnancy.
surfactant secretion after differentiation. They further This breakthrough offers new possibilities for prenatal
cultured lung organoids from the AF and tracheal fluid diagnosis and treatment of congenital diseases.

Volume 1 Issue 2 (2025) 5 doi: 10.36922/OR025040005