Human menstrual blood-derived stem cells (hMenSCs) show promise in regenerative medicine and endometrial disease treatments. Organoids, three-dimensional (3D) tissue replicas, provide accurate disease models. Fibroblast growth factor 2 (FGF2), with established roles in stem cell functions, remains underexplored in its impact on hMenSCs and endometrial organoids. Our study aims to explore these unexplored effects of FGF2. In our study, we have developed a method for rapid and high-quality cultivation of MenSCs and organoids. Using this novel platform, we demonstrate that FGF2 alters hMenSC morphology, enhancing growth and proliferative, migratory, and anti-aging capabilities. RNA sequencing suggests this is linked to downregulated cell adhesion genes. FGF2 promotes proliferation via ribosomal biogenesis and ATP metabolism, possibly delaying senescence. In organoids, FGF2 reverses compaction and volume reduction, promoting growth and fibroblast release. q-PCR analysis shows FGF2 regulates genes related to stemness, proliferation, metabolism, and PI3K-AKT signaling, with distinct patterns in hMenSCs and organoids. This research advances regenerative medicine and reproductive biology by optimizing the cultivation of hMenSCs and organoids with FGF2 for potential therapeutic applications.