Comparative transcriptomic profiling of human 3D endometrial stromal organoids under distinct decidualization stimuli

Three-dimensional (3D) endometrial organoid systems provide a physiologically relevant platform to study this process; however, how different stimulatory conditions influence decidualization responses in 3D culture remains incompletely characterized. In this study, we generated 3D human endometrial stromal organoids and subjected them to four experimental conditions: vehicle control, cyclic adenosine monophosphate (cAMP) stimulation, combined estrogen–progesterone–cAMP (EPC) treatment, and co-culture with in vitro–grown follicles derived from mouse ovary.

Organoids were cultured for 6 days under each condition, followed by bulk RNA sequencing. Principal component analysis (PCA) revealed clear separation between control and stimulated groups, with cAMP, EPC, and follicle co-culture conditions showing distinct but partially overlapping transcriptional profiles. Differential gene expression analysis identified both shared and condition-specific transcriptional changes across the four treatments.