Three-dimensional cell cultures as an in vitro tool for ovarian cancer modeling and natural product discovery

Ovarian cancer (OC) remains the most common cancer among women, with high incidence and mortality rates, prompting extensive investigation across in vivo, in vitro, and clinical settings. In this context, in vitro cell culture models have been widely used to study drug response, tumor progression, and the development of novel therapeutic strategies. Among these, three-dimensional (3D) culture systems have emerged as advanced platforms capable of more accurately recapitulating tumor complexity. Concurrently, the exploration of complementary therapeutic approaches, especially natural products, has gained attention for their potential to enhance antitumoral efficacy while reducing adverse effects. In this interpretative narrative review, we examine the application of 3D culture models to evaluate the effects of natural compounds in OC. Based on 21 selected studies, we highlight that 3D systems, including spheroid formation via droplet suspension, ultra-low adhesion plates, coated surfaces, hydrogels, and rotary culture, enable more physiologically relevant modeling of the tumor microenvironment. Within these systems, a wide range of natural compounds (e.g., epigallocatechin gallate, withaferin A, lycopene, cucurbitacins, genistein, and others) consistently exhibited antitumor activity. These effects include inhibition of migration and invasion, induction of apoptosis, modulation of oxidative and inflammatory pathways, and disruption of cell-matrix interactions. Mechanistically, these natural compounds target key signaling pathways involved in tumor progression, although significant gaps remain regarding standardization, reproducibility, and clinical translation. Importantly, this review underscores the superiority of 3D in vitro models in recapitulating tumor biology and highlights the promising role of natural products as adjuvant strategies alongside conventional therapies. Collectively, these findings provide a robust framework for advancing experimental, translational, and clinical research aimed at improving therapeutic outcomes in OC.