Simple and Rapid Model to Generate Differentiated Endometrial Floating Organoids

droplets in the presence of standard culture medium. From a structural point of view, these novel floating 3D cultures develop as gland-like structures constituted by epithelial cells organized around a central lumen and retain the expression of endometrial and decidual genes, like previously published organoids, although with a phenotype resembling hormonally differentiated structures. Importantly, floating organoids retain stromal cells which grow in close contact with the epithelial cells, localized within the internal or external portion of the organoid structure. In summary, we present a simple and rapid model for generating 3D endometrial organoids that preserve epithelial-stromal cell interactions, promoting the formation of differentiated organoids and enabling the study of reciprocal modulation between epithelium and stroma. Key features • Development of 3D endometrial organoids using 10% FBS-DMEM/F12 complete medium in less than a month. • Preservation of stromal and epithelial cell populations, favoring the study of their interaction. • Development of differentiated endometrial organoids without the need for in vitro hormonal stimulation.