Formation of an endometrial epithelial monolayer in a microfluidic device with human tissue-derived endometrial organoids

The endometrium is the uterine lining that supports implantation and pregnancy. Existing <i>in vitro</i> systems only partly capture epithelial structure and function. We built a microfluidic model of the human endometrial epithelium using patient-derived organoids and defined a parameterized device and ECM conditions that yield a stable, polarized monolayer on chip. We specify the geometry, surface treatments, and collagen-based hydrogel or coating conditions, and we link these parameters to epithelial morphology and barrier integrity readouts. The epithelial layer maintains histologic features and endometrium-relevant markers and shows hormone-responsive transcript profiles. We quantify donor-to-donor variability across two donors and use it as a design constraint for reproducible culture. Because stromal and immune components shape the reproductive microenvironment, we will extend this platform to modular multicellular co-cultures that incorporate these elements.