Zhou, JSheridan, MATian, YDahlgren, KJMessler, MPeng, TZhao, AEzashi, TSchulz, LCUlery, BDRoberts, RMSchust, DJ2025-06-172025-06-172025-032589-00422589-0042https://hdl.handle.net/10161/32494The development of trophoblast organoids has enabled investigation of placental physiology, disease, and early maternal-fetal interactions during a previously restricted stage of pregnancy. A key shortcoming in existing trophoblast organoid methodologies is the non-physiologic position of the syncytiotrophoblast (STB) within the inner portion of the organoid, which neither recapitulates <i>in vivo</i> placental villous morphology nor allows for facile modeling of STB exposure to the endometrium or the contents of the intervillous space. Here, we have successfully established apical-out human trophoblast stem cells (hTSC)-sourced organoids with STB forming on the surface of the organoid. These organoids can also be induced to give rise to the extravillous trophoblast (EVT) lineage, which invades into an extracellular matrix-based hydrogel. Compared to previous methods, our organoids more closely mimic developing human placental architecture, offering a novel platform to study normal and abnormal placental development and to model exposures to pharmaceuticals, pathogens, and environmental factors.https://creativecommons.org/licenses/by-nc/4.0Cell biologyMolecular biologyPhysiologyJournal article