Hydrogel-Based Three-Dimensional Culture Systems in Reproductive Technology: A Review

Abstract

Context: Assisted reproductive technologies (ARTs), particularly in vitro embryo production (IVEP), are important tools for improving reproductive efficiency and accelerating genetic progress. However, conventional two-dimensional (2D) culture systems have major limitations because they cannot fully recapitulate the complex physiological microenvironment of ovarian follicles. This limitation may disrupt cellular communication, impair gap junctions, reduce oocyte developmental competence, and ultimately result in poor-quality embryos. This review examines the shift from 2D to three-dimensional (3D) culture systems in reproductive technology, with a specific focus on hydrogel-based scaffolds that mimic the natural extracellular matrix (ECM) and provide a more physiological environment for cell growth and interactions. Evidence Acquisition: This narrative review analyzed studies evaluating hydrogel-based 3D culture systems in reproductive applications. The review focused on evidence regarding how these systems affect follicle survival and function, oocyte quality, and embryonic developmental potential across species. The available evidence indicates that 3D systems can preserve follicular and embryonic architecture, enhance nutrient and gas exchange, and restore essential cell-to-cell and cell-matrix interactions. Results: Evidence indicates that hydrogel-based scaffolds, by preserving three-dimensional architecture and restoring essential cellular interactions, can improve in vitro maturation (IVM) rates, embryonic developmental competence, and overall IVF outcomes. Natural hydrogels, including ECM-based, alginate-based, agarose-based, hyaluronic acid-based, and chitosan-based scaffolds, as well as synthetic systems such as polyethylene glycol (PEG)-based hydrogels, have demonstrated promising outcomes in follicle culture, oocyte maturation, and embryo development. However, species-specific responses and differences in scaffold concentration, stiffness, biochemical composition, and degradation behavior remain key determinants of success. Conclusions: Hydrogel-based 3D culture systems offer a promising strategy for improving reproductive technologies by more closely simulating in vivo conditions. Their future application in reproductive medicine, fertility preservation, and animal biotechnology will depend on further optimization of scaffold composition, mechanical properties, biodegradability, and species-specific culture requirements.