Ex vivo model for assessing fetal membrane integrity and therapeutic strategies

The fetal membranes (FM) are essential for maintaining the fetal–maternal interface and ensuring mechanical and barrier integrity throughout pregnancy. Preterm prelabor rupture of FM (pPROM) and iatrogenic pPROM (iPPROM) following fetal surgery, remain major causes of preterm birth. To better understand potential FM repair mechanisms and develop preventive strategies, experimental models that remain stable and functional beyond only a few days of culture are particularly needed. Here, we present a modular ex vivo model that enables up to 21 days culture of full-thickness human FM. Quantification of DNA content, ATP levels, glucose consumption, and lactate production suggest sustained but gradually adapting metabolic activity. The largely intact epithelial and mesenchymal organization and FM barrier function reveal maintained overall structural integrity. Additionally, mechanical testing demonstrates highest FM rupture pressure resistance close to the placental region with a progressive decrease with increasing distance from the placenta and preserved mechanical stability after 14 days of culture. Finally, puncture experiments show a time-dependent collagen fiber realignment around puncture-induced FM defects. Together, this accessible and adaptable platform provides a human-based experimental toolbox for investigating FM biology, evaluating biomaterials, and studying defect healing in a clinically relevant context. By enabling extended culture and integrated functional readouts, the model may help to bridge the translational gap between short-term in vitro systems and in vivo studies in maternal–fetal medicine.