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Although models such as gland\u2010like structures (GLSs) and endometrial assembloids (EnAos) are successfully established, they lack an intact luminal epithelium, which makes it difficult to recapitulate endometrial receptivity. Here, a novel EnAo model (ALI\u2010EnAo) is developed by combining endometrial epithelial cells (EnECs) and stromal cells (EnSCs) and using an improved matrix and air\u2013liquid interface (ALI) culture method. ALI\u2010EnAos exhibit intact EnSCs and glandular and luminal epithelia, which recapitulates human endometrium anatomy, cell composition, hormone\u2010induced menstrual cycle changes, gene expression profiles, and dynamic ciliogenesis. The model suggests that EnSCs, together with the extracellular matrix and ALI culture conditions, contribute to EnAo phenotypes and characteristics reflective of the endometrial menstrual cycle. This enables to transcriptionally define endometrial cell subpopulations. It anticipates that ALI\u2010EnAos will facilitate studies on embryo implantation, and endometrial growth, differentiation, and disease.<\/jats:p>","DOI":"10.1002\/advs.202301868","type":"journal-article","created":{"date-parts":[[2023,8,27]],"date-time":"2023-08-27T22:58:21Z","timestamp":1693177101000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":48,"title":["Generation of Human Endometrial Assembloids with a Luminal Epithelium using Air\u2013Liquid Interface Culture Methods"],"prefix":"10.1002","volume":"10","author":[{"given":"Jiwen","family":"Tian","sequence":"first","affiliation":[{"name":"State Key Laboratory of Primate Biomedical Research Institute of Primate Translational Medicine Kunming University of Science and Technology  Kunming Yunnan 650032 China"},{"name":"Medical School Kunming University of Science and Technology  Kunming Yunnan 650032 China"},{"name":"Department of Reproductive Medicine The First People's 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